Methods of preparing inhibitors of influenza viruses replication

ABSTRACT

A method of preparing a compound of Formula (I) or a pharmaceutically acceptable salt thereof: 
     
       
         
         
             
             
         
       
     
     comprises: (a) reacting Compound (1): 
     
       
         
         
             
             
         
       
     
     or a pharmaceutically acceptable salt thereof with Compound (2): 
     
       
         
         
             
             
         
       
     
     in the presence of water, an organic solvent, a base, and a transition metal catalyst to generate a compound of Formula (I) or a pharmaceutically acceptable salt thereof.

CROSS REFERENCE TO RELATED APPLICATION

This PCT application claims the benefit of U.S. provisional applicationNo. 61/160,636, filed on May 13, 2015. This document is herebyincorporated by reference in its entirety.

FIELD OF THE INVENTION

The present invention relates to processes and intermediates useful forthe preparation of compounds that possess inhibitory activity forInfluenza virus replication.

BACKGROUND OF THE INVENTION

Influenza spreads around the world in seasonal epidemics, resulting inthe deaths of hundreds of thousands annually—millions in pandemic years.For example, three influenza pandemics occurred in the 20th century andkilled tens of millions of people, with each of these pandemics beingcaused by the appearance of a new strain of the virus in humans. Often,these new strains result from the spread of an existing influenza virusto humans from other animal species.

Influenza is primarily transmitted from person to person via largevirus-laden droplets that are generated when infected persons cough orsneeze; these large droplets can then settle on the mucosal surfaces ofthe upper respiratory tracts of susceptible individuals who are near(e.g. within 6 feet) infected persons. Transmission might also occurthrough direct contact or indirect contact with respiratory secretions,such as touching surfaces contaminated with influenza virus and thentouching the eyes, nose or mouth. Adults might be able to spreadinfluenza to others from 1 day before getting symptoms to approximately5 days after symptoms start. Young children and persons with weakenedimmune systems might be infectious for 10 or more days after onset ofsymptoms.

Influenza viruses are RNA viruses of the family Orthomyxoviridae, whichcomprises five genera: Influenza virus A, Influenza virus B, Influenzavirus C, ISA virus and Thogoto virus.

The Influenza virus A genus has one species, influenza A virus. Wildaquatic birds are the natural hosts for a large variety of influenza A.Occasionally, viruses are transmitted to other species and may thencause devastating outbreaks in domestic poultry or give rise to humaninfluenza pandemics. The type A viruses are the most virulent humanpathogens among the three influenza types and cause the most severedisease. The influenza A virus can be subdivided into differentserotypes based on the antibody response to these viruses. The serotypesthat have been confirmed in humans, ordered by the number of known humanpandemic deaths, are: H1N1 (which caused Spanish influenza in 1918),H2N2 (which caused Asian Influenza in 1957), H3N2 (which caused HongKong Flu in 1968), H5N1 (a pandemic threat in the 2007-2008 influenzaseason), H7N7 (which has unusual zoonotic potential), H1N2 (endemic inhumans and pigs), H9N2, H7N2, H7N3 and H10N7.

The Influenza virus B genus has one species, influenza B virus.Influenza B almost exclusively infects humans and is less common thaninfluenza A. The only other animal known to be susceptible to influenzaB infection is the seal. This type of influenza mutates at a rate 2-3times slower than type A and consequently is less genetically diverse,with only one influenza B serotype. As a result of this lack ofantigenic diversity, a degree of immunity to influenza B is usuallyacquired at an early age. However, influenza B mutates enough thatlasting immunity is not possible. This reduced rate of antigenic change,combined with its limited host range (inhibiting cross species antigenicshift), ensures that pandemics of influenza B do not occur.

The Influenza virus C genus has one species, influenza C virus, whichinfects humans and pigs and can cause severe illness and localepidemics. However, influenza C is less common than the other types andusually seems to cause mild disease in children.

Influenza A, B and C viruses are very similar in structure. The virusparticle is 80-120 nanometers in diameter and usually roughly spherical,although filamentous forms can occur. Unusually for a virus, its genomeis not a single piece of nucleic acid; instead, it contains seven oreight pieces of segmented negative-sense RNA. The Influenza A genomeencodes 11 proteins: hemagglutinin (HA), neuraminidase (NA),nucleoprotein (NP), M1, M2, NS1, NS2(NEP), PA, PB1, PB1-F2 and PB2.

HA and NA are large glycoproteins on the outside of the viral particles.HA is a lectin that mediates binding of the virus to target cells andentry of the viral genome into the target cell, while NA is involved inthe release of progeny virus from infected cells, by cleaving sugarsthat bind the mature viral particles. Thus, these proteins have beentargets for antiviral drugs. Furthermore, they are antigens to whichantibodies can be raised. Influenza A viruses are classified intosubtypes based on antibody responses to HA and NA, forming the basis ofthe H and N distinctions (vide supra) in, for example, H5N1.

Influenza produces direct costs due to lost productivity and associatedmedical treatment, as well as indirect costs of preventative measures.In the United States, influenza is responsible for a total cost of over$10 billion per year, while it has been estimated that a future pandemiccould cause hundreds of billions of dollars in direct and indirectcosts. Preventative costs are also high. Governments worldwide havespent billions of U.S. dollars preparing and planning for a potentialH5N1 avian influenza pandemic, with costs associated with purchasingdrugs and vaccines as well as developing disaster drills and strategiesfor improved border controls.

Current treatment options for influenza include vaccination, andchemotherapy or chemoprophylaxis with anti-viral medications.Vaccination against influenza with an influenza vaccine is oftenrecommended for high-risk groups, such as children and the elderly, orin people that have asthma, diabetes, or heart disease. However, it ispossible to get vaccinated and still get influenza. The vaccine isreformulated each season for a few specific influenza strains but cannotpossibly include all the strains actively infecting people in the worldfor that season. It may take six months for the manufacturers toformulate and produce the millions of doses required to deal with theseasonal epidemics; occasionally, a new or overlooked strain becomesprominent during that time and infects people although they have beenvaccinated (as by the H3N2 Fujian flu in the 2003-2004 influenzaseason). It is also possible to get infected just before vaccination andget sick with the very strain that the vaccine is supposed to prevent,as the vaccine may take several weeks to become effective.

Further, the effectiveness of these influenza vaccines is variable. Dueto the high mutation rate of the virus, a particular influenza vaccineusually confers protection for no more than a few years. A vaccineformulated for one year may be ineffective in the following year, sincethe influenza virus changes rapidly over time, and different strainsbecome dominant.

Also, because of the absence of RNA proofreading enzymes, theRNA-dependent RNA polymerase of influenza vRNA makes a single nucleotideinsertion error roughly every 10 thousand nucleotides, which is theapproximate length of the influenza vRNA. Hence, nearly everynewly-manufactured influenza virus is a mutant-antigenic drift. Theseparation of the genome into eight separate segments of vRNA allowsmixing or reassortment of vRNAs if more than one viral line has infecteda single cell. The resulting rapid change in viral genetics producesantigenic shifts and allows the virus to infect new host species andquickly overcome protective immunity.

Antiviral drugs can also be used to treat influenza, with neuraminidaseinhibitors being particularly effective, but viruses can developresistance to the standard antiviral drugs.

Thus, there is still a need for drugs for treating influenza infections,such as for drugs with expanded treatment window, and/or reducedsensitivity to viral titer. Further, there is a need for methods forpreparing such drugs efficiently.

SUMMARY OF THE INVENTION

The present invention generally relates to methods of preparing acompound of Formula (I),

or a pharmaceutically acceptable salt thereof, wherein R¹ is —Cl or —F,and to methods of preparing certain intermediate compounds therefor.

In one embodiment, the invention is directed to a process for preparinga compound of Formula I:

or a pharmaceutically acceptable salt thereof, wherein R¹ is —Cl or —F;comprising the step of:

i) reacting a compound of Formula 1 with a compound of Formula 2

in the presence of water, an organic solvent, a base, and a transitionmetal catalyst to generate a compound of Formula I; wherein each of R⁸and R⁹ is —OH, —O—C₁₋₄ aliphatic optionally substituted with 1-5occurrences of R¹¹, or R⁸ and R⁹ together with the boron atom to whichthey are attached, form a 5-9 membered mono- or bicyclic ring system,optionally substituted with 1-6 occurrences of R¹¹, or BF₃K; each R¹¹ isindependently selected from halogen, —OCH₃, —OH, —NO₂, —NH₂, —SH, —SCH₃,—NHCH₃, —CN, ═O, or unsubstituted —C₁₋₂ aliphatic; X¹ is —Cl, —Br, —I,—OTs, —OMs, —OH, or —NH₂; andthe transition metal catalyst is

or the transition metal catalyst comprises Pd and a ligand comprising

Some embodiments further comprise the step of:

ii) deprotecting a compound of Formula 3 to generate a compound ofFormula I:

In some embodiments, step ii) comprises deprotecting the compound ofFormula 3 in the presence of a base. In some examples, the basecomprises an inorganic base. For instance, the inorganic base is analkali metal hydroxide. In other examples, the alkali metal hydroxide isLiOH, NaOH, KOH, or any combination thereof.

In another embodiment, the invention is directed to a process forpreparing a compound of Formula I:

or a pharmaceutically acceptable salt thereof, wherein R¹ is —Cl or —F;comprising the step of:

i) reacting a compound of Formula 1 with a compound of Formula 2

in the presence of water, an organic solvent, a base, and a transitionmetal catalyst to generate a compound of Formula 3; and

ii) deprotecting a compound of Formula 3 to generate a compound ofFormula I:

wherein each of R⁸ and R⁹ is —OH, —O—C₁₋₄ aliphatic optionallysubstituted with 1-5 occurrences of R¹¹, or R⁸ and R⁹ together with theboron atom to which they are attached, form a 5-9 membered mono- orbicyclic ring system, optionally substituted with 1-6 occurrences ofR¹¹, or BF₃K; each R¹¹ is independently selected from halogen, —OCH₃,—OH, —NO₂, —NH₂, —SH, —SCH₃, —NHCH₃, —CN, ═O, or unsubstituted —C₁₋₂aliphatic; X¹ is —Cl, —Br, —I, —OTs, —OMs, —OH, or —NH₂; andthe transition metal catalyst is

or the transition metal catalyst comprises Pd and a ligand comprising

In some embodiments, the base of step i) is an organic base. Forexample, the organic base comprises a tertiary amine. And, in someinstances, the tertiary amine comprises diisopropylethylamine,triethylamine, triethylenediamine, or any combination thereof.

In some embodiments, the organic solvent of step i) is an aproticsolvent. For example, the aprotic solvent is acetonitrile, toluene,N,N-dimethylformamide, N,N-dimethylacetamide, acetone, methyl tert-butylether, or any combination thereof.

In some embodiments, R¹ is —F. In other embodiments, R¹ is —Cl.

In some embodiments, X¹ is —Cl.

In some embodiments, the reaction of step i) is performed at atemperature between about 50° C. and about 110° C. For example, thereaction of step i) is performed at a temperature between about 60° C.and about 95° C. In other examples, the reaction of step i) is performedat a temperature between about 70° C. and about 80° C.

Some embodiments further comprising the step of:

via) reacting a compound of Formula 8 with a compound of Formula 9,

in the presence of a base and an organic solvent to generate a mixturecomprising a compound of Formula 2 and a compound of Formula 10:

Some embodiments further comprise the steps of:

vii) reacting the mixture comprising the compound of Formula 2 and thecompound of Formula 10 with HCl in the presence of an organic solvent togenerate a mixture of hydrochloride salts of the compound of Formula 2and the compound of Formula 10; and

viii) recrystalizing the mixture of the hydrochloride salts of thecompound of Formula 2 and the compound of Formula 10 to generate thehydrochloride salt of the compound of Formula 2.

Some embodiments further comprising the steps of:

vib) reacting a compound of Formula 8 with an acid salt of a compound ofFormula 9 in the presence of a solvent and a base to generate thecompound Formula 2

and

viib) reacting the compound of Formula 2 with HCl to generate thehydrochloride salt of the compound of Formula 2.

In some embodiments, the base of step vib) is an inorganic base selectedfrom tripotassium phosphate, dipotassium hydrogen phosphate, dipotassiumcarbonate, disodium carbonate, trisodium phosphate, disodium hydrogenphosphate, or any combination thereof.

In some embodiments, the solvent of step vib) comprises water.

In some embodiments, the solvent of step vib) further comprises analcohol selected from methanol, ethanol, propanol, iso-propanol,butanol, tert-butanol, or any combination thereof.

In some embodiments, the reaction of step vib) is performed at atemperature of from about 50° C. to about 100° C. For example, thereaction of step vib) is performed at a temperature of from about 60° C.to about 80° C.

In some embodiments, the compound of Formula 1 is 1a

In some embodiments, the compound of Formula 2 is 2a

In some embodiments, the compound of Formula I is Ia

DETAILED DESCRIPTION OF THE INVENTION

The present invention provides methods and intermediates forsynthesizing compounds that possess inhibitory activity for influenzavirus replication.

I. COMMONLY USED ABBREVIATIONS

-   -   ACN acetonitrile    -   tBuOAc tert-butyl acetate    -   DABCO 1,4-diazabicyclo[2.2.2]octane    -   DCM dichloromethane    -   EtOAc ethyl acetate    -   IPAc iso-propyl acetate    -   MIBK methyl iso-butyl ketone    -   TEA triethylamine    -   THF tetrahydrofuran    -   PG protecting group    -   LG leaving group    -   Ac acetyl    -   TMS trimethylsilyl    -   TBS tert-butyldimethylsilyl    -   TIPS tri-iso-propylsilyl    -   TBDPS tert-butyldiphenylsilyl    -   TOM tri-iso-propylsilyloxymethyl    -   DMP Dess-Martin periodinane    -   IBX 2-iodoxybenzoic acid    -   DMF dimethylformamide    -   MTBE methyl-tert-butylether    -   TBAF tetra-n-butylammonium fluoride    -   d.e. diastereomeric excess    -   e.e. enantiomeric excess    -   d.r. diastereomeric ratio    -   DMSO dimethyl sulfoxide    -   TCA trichloroacetic acid    -   ATP adenosine triphosphate    -   EtOH ethanol    -   Ph phenyl    -   Me methyl    -   Et ethyl    -   Bu butyl    -   DEAD diethylazodicarboxylate    -   HEPES 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid    -   DTT dithiothreitol    -   MOPS 4-morpholinepropanesulfonic acid    -   NMR nuclear magnetic resonance    -   HPLC high performance liquid chromatography    -   LCMS liquid chromatography-mass spectrometry    -   TLC thin layer chromatography    -   Rt retention time    -   HOBt hydroxybenzotriazole    -   Ms mesyl    -   Ts tosyl    -   Tf triflyl    -   Bs besyl    -   Ns nosyl    -   Cbz carboxybenzyl    -   Moz p-methoxybenzyl carbonyl    -   Boc tert-butyloxycarbonyl    -   Fmoc 9-fluorenylmethyloxycarbonyl    -   Bz benzoyl    -   Bn benzyl    -   PMB p-methoxybenzyl    -   AUC area under the curve    -   DMPM 3,4-dimethoxybenzyl    -   PMP p-methoxyphenyl    -   XRPD X-ray powder diffraction    -   CbzOSu N-(benzyloxycarbonyloxy)succinimide    -   RRT relative retention time    -   DIPEA N,N-diisopropylethylamine    -   dppf 1,1′-bis(diphenylphosphino)ferrocene    -   CHex cyclohexane    -   dba dibenzylideneacetone

II. DEFINITIONS

For purposes of this invention, the chemical elements are identified inaccordance with the Periodic Table of the Elements, CAS version,Handbook of Chemistry and Physics, 75th Ed. Additionally, generalprinciples of organic chemistry are described in “Organic Chemistry”,Thomas Sorrell, University Science Books, Sausalito: 1999, and “March'sAdvanced Organic Chemistry”, 5th Ed., Ed.: Smith, M. B. and March, J.,John Wiley & Sons, New York: 2001, the entire contents of which arehereby incorporated by reference.

As described herein, compounds of the invention may optionally besubstituted with one or more substituents, such as are illustratedgenerally above, or as exemplified by particular classes, subclasses,and species of the invention.

As used herein, the term “hydroxyl” or “hydroxy” refers to an —OHmoiety.

As used herein the term “aliphatic” encompasses the terms alkyl,alkenyl, alkynyl, each of which being optionally substituted as setforth below.

As used herein, an “alkyl” group refers to a saturated aliphatichydrocarbon group containing 1-12 (e.g., 1-8, 1-6, or 1-4) carbon atoms.An alkyl group can be straight or branched. Examples of alkyl groupsinclude, but are not limited to, methyl, ethyl, propyl, isopropyl,butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, n-heptyl, or2-ethylhexyl. An alkyl group can be substituted (i.e., optionallysubstituted) with one or more substituents such as halo, phospho,cycloaliphatic [e.g., cycloalkyl or cycloalkenyl], heterocycloaliphatic[e.g., heterocycloalkyl or heterocycloalkenyl], aryl, heteroaryl,alkoxy, aroyl, heteroaroyl, acyl [e.g., (aliphatic)carbonyl,(cycloaliphatic)carbonyl, or (heterocycloaliphatic)carbonyl], nitro,cyano, amido [e.g., (cycloalkylalkyl)carbonylamino, arylcarbonylamino,aralkylcarbonylamino, (heterocycloalkyl)carbonylamino,(heterocycloalkylalkyl)carbonylamino, heteroarylcarbonylamino,heteroaralkylcarbonylamino alkylaminocarbonyl, cycloalkylaminocarbonyl,heterocycloalkylaminocarbonyl, arylaminocarbonyl, orheteroarylaminocarbonyl], amino [e.g., aliphaticamino,cycloaliphaticamino, or heterocycloaliphaticamino], sulfonyl [e.g.,aliphatic-SO₂—], sulfinyl, sulfanyl, sulfoxy, urea, thiourea, sulfamoyl,sulfamide, oxo, carboxy, carbamoyl, cycloaliphaticoxy,heterocycloaliphaticoxy, aryloxy, heteroaryloxy, aralkyloxy,heteroarylalkoxy, alkoxycarbonyl, alkylcarbonyloxy, or hydroxy. Withoutlimitation, some examples of substituted alkyls include carboxyalkyl(such as HOOC-alkyl, alkoxycarbonylalkyl, and alkylcarbonyloxyalkyl),cyanoalkyl, hydroxyalkyl, alkoxyalkyl, acylalkyl, aralkyl,(alkoxyaryl)alkyl, (sulfonylamino)alkyl (such as(alkyl-SO₂-amino)alkyl), aminoalkyl, amidoalkyl, (cycloaliphatic)alkyl,or haloalkyl.

As used herein, an “alkenyl” group refers to an aliphatic carbon groupthat contains 2-8 (e.g., 2-12, 2-6, or 2-4) carbon atoms and at leastone double bond. Like an alkyl group, an alkenyl group can be straightor branched. Examples of an alkenyl group include, but are not limitedto allyl, 1- or 2-isopropenyl, 2-butenyl, and 2-hexenyl. An alkenylgroup can be optionally substituted with one or more substituents suchas halo, phospho, cycloaliphatic [e.g., cycloalkyl or cycloalkenyl],heterocycloaliphatic [e.g., heterocycloalkyl or heterocycloalkenyl],aryl, heteroaryl, alkoxy, aroyl, heteroaroyl, acyl [e.g.,(aliphatic)carbonyl, (cycloaliphatic)carbonyl, or(heterocycloaliphatic)carbonyl], nitro, cyano, amido [e.g.,(cycloalkylalkyl)carbonylamino, arylcarbonylamino, aralkylcarbonylamino,(heterocycloalkyl)carbonylamino, (heterocycloalkylalkyl)carbonylamino,heteroarylcarbonylamino, heteroaralkylcarbonylamino alkylaminocarbonyl,cycloalkylaminocarbonyl, heterocycloalkylaminocarbonyl,arylaminocarbonyl, or heteroarylaminocarbonyl], amino [e.g.,aliphaticamino, cycloaliphaticamino, heterocycloaliphaticamino, oraliphaticsulfonylamino], sulfonyl [e.g., alkyl-SO₂—,cycloaliphatic-SO₂—, or aryl-SO₂—], sulfinyl, sulfanyl, sulfoxy, urea,thiourea, sulfamoyl, sulfamide, oxo, carboxy, carbamoyl,cycloaliphaticoxy, heterocycloaliphaticoxy, aryloxy, heteroaryloxy,aralkyloxy, heteroaralkoxy, alkoxycarbonyl, alkylcarbonyloxy, orhydroxy. Without limitation, some examples of substituted alkenylsinclude cyanoalkenyl, alkoxyalkenyl, acylalkenyl, hydroxyalkenyl,aralkenyl, (alkoxyaryl)alkenyl, (sulfonylamino)alkenyl (such as(alkyl-SO₂-amino)alkenyl), aminoalkenyl, amidoalkenyl,(cycloaliphatic)alkenyl, or haloalkenyl.

As used herein, an “alkynyl” group refers to an aliphatic carbon groupthat contains 2-8 (e.g., 2-12, 2-6, or 2-4) carbon atoms and has atleast one triple bond. An alkynyl group can be straight or branched.Examples of an alkynyl group include, but are not limited to, propargyland butynyl. An alkynyl group can be optionally substituted with one ormore substituents such as aroyl, heteroaroyl, alkoxy, cycloalkyloxy,heterocycloalkyloxy, aryloxy, heteroaryloxy, aralkyloxy, nitro, carboxy,cyano, halo, hydroxy, sulfo, mercapto, sulfanyl [e.g., aliphaticsulfanylor cycloaliphaticsulfanyl], sulfinyl [e.g., aliphaticsulfinyl orcycloaliphaticsulfinyl], sulfonyl [e.g., aliphatic-SO₂—,aliphaticamino-SO₂—, or cycloaliphatic-SO₂—], amido [e.g.,aminocarbonyl, alkylaminocarbonyl, alkylcarbonylamino,cycloalkylaminocarbonyl, heterocycloalkylaminocarbonyl,cycloalkylcarbonylamino, arylaminocarbonyl, arylcarbonylamino,aralkylcarbonylamino, (heterocycloalkyl)carbonylamino,(cycloalkylalkyl)carbonylamino, heteroaralkylcarbonylamino,heteroarylcarbonylamino or heteroarylaminocarbonyl], urea, thiourea,sulfamoyl, sulfamide, alkoxycarbonyl, alkylcarbonyloxy, cycloaliphatic,heterocycloaliphatic, aryl, heteroaryl, acyl [e.g.,(cycloaliphatic)carbonyl or (heterocycloaliphatic)carbonyl], amino[e.g., aliphaticamino], sulfoxy, oxo, carboxy, carbamoyl,(cycloaliphatic)oxy, (heterocycloaliphatic)oxy, or (heteroaryl)alkoxy.

As used herein, an “amido” encompasses both “aminocarbonyl” and“carbonylamino”. These terms when used alone or in connection withanother group refer to an amido group such as —N(R^(X))—C(O)—R^(Y) or—C(O)—N(R^(X))₂, when used terminally, and —C(O)—N(R^(X))— or—N(R^(X))—C(O)— when used internally, wherein R^(X) and R^(Y) can bealiphatic, cycloaliphatic, aryl, araliphatic, heterocycloaliphatic,heteroaryl or heteroaraliphatic. Examples of amido groups includealkylamido (such as alkylcarbonylamino or alkylaminocarbonyl),(heterocycloaliphatic)amido, (heteroaralkyl)amido, (heteroaryl)amido,(heterocycloalkyl)alkylamido, arylamido, aralkylamido,(cycloalkyl)alkylamido, or cycloalkylamido.

As used herein, an “amino” group refers to —NR^(X)R^(Y) wherein each ofR^(X) and R^(Y) is independently hydrogen, aliphatic, cycloaliphatic,(cycloaliphatic)aliphatic, aryl, araliphatic, heterocycloaliphatic,(heterocycloaliphatic)aliphatic, heteroaryl, carboxy, sulfanyl,sulfinyl, sulfonyl, (aliphatic)carbonyl, (cycloaliphatic)carbonyl,((cycloaliphatic)aliphatic)carbonyl, arylcarbonyl,(araliphatic)carbonyl, (heterocycloaliphatic)carbonyl,((heterocycloaliphatic)aliphatic)carbonyl, (heteroaryl)carbonyl, or(heteroaraliphatic)carbonyl, each of which being defined herein andbeing optionally substituted. Examples of amino groups includealkylamino, dialkylamino, or arylamino. When the term “amino” is not theterminal group (e.g., alkylcarbonylamino), it is represented by—NR^(X)—, where R^(X) has the same meaning as defined above.

As used herein, an “aryl” group used alone or as part of a larger moietyas in “aralkyl”, “aralkoxy”, or “aryloxyalkyl” refers to monocyclic(e.g., phenyl); bicyclic (e.g., indenyl, naphthalenyl,tetrahydronaphthyl, tetrahydroindenyl); and tricyclic (e.g., fluorenyltetrahydrofluorenyl, or tetrahydroanthracenyl, anthracenyl) ring systemsin which the monocyclic ring system is aromatic or at least one of therings in a bicyclic or tricyclic ring system is aromatic. The bicyclicand tricyclic groups include benzofused 2-3 membered carbocyclic rings.For example, a benzofused group includes phenyl fused with two or moreC₄₋₈ carbocyclic moieties. An aryl is optionally substituted with one ormore substituents including aliphatic [e.g., alkyl, alkenyl, oralkynyl]; cycloaliphatic; (cycloaliphatic)aliphatic;heterocycloaliphatic; (heterocycloaliphatic)aliphatic; aryl; heteroaryl;alkoxy; (cycloaliphatic)oxy; (heterocycloaliphatic)oxy; aryloxy;heteroaryloxy; (araliphatic)oxy; (heteroaraliphatic)oxy; aroyl;heteroaroyl; amino; oxo (on a non-aromatic carbocyclic ring of abenzofused bicyclic or tricyclic aryl); nitro; carboxy; amido; acyl[e.g., (aliphatic)carbonyl; (cycloaliphatic)carbonyl;((cycloaliphatic)aliphatic)carbonyl; (araliphatic)carbonyl;(heterocycloaliphatic)carbonyl;((heterocycloaliphatic)aliphatic)carbonyl; or(heteroaraliphatic)carbonyl]; sulfonyl [e.g., aliphatic-SO₂— oramino-SO₂—]; sulfinyl [e.g., aliphatic-S(O)— or cycloaliphatic-S(O)—];sulfanyl [e.g., aliphatic-S—]; cyano; halo; hydroxy; mercapto; sulfoxy;urea; thiourea; sulfamoyl; sulfamide; or carbamoyl. Alternatively, anaryl can be unsubstituted.

Non-limiting examples of substituted aryls include haloaryl [e.g.,mono-, di (such as p,m-dihaloaryl), and (trihalo)aryl]; (carboxy)aryl[e.g., (alkoxycarbonyl)aryl, ((aralkyl)carbonyloxy)aryl, and(alkoxycarbonyl)aryl]; (amido)aryl [e.g., (aminocarbonyl)aryl,(((alkylamino)alkyl)aminocarbonyl)aryl, (alkylcarbonyl)aminoaryl,(arylaminocarbonyl)aryl, and (((heteroaryl)amino)carbonyl)aryl];aminoaryl [e.g., ((alkylsulfonyl)amino)aryl or ((dialkyl)amino)aryl];(cyanoalkyl)aryl; (alkoxy)aryl; (sulfamoyl)aryl [e.g.,(aminosulfonyl)aryl]; (alkylsulfonyl)aryl; (cyano)aryl;(hydroxyalkyl)aryl; ((alkoxy)alkyl)aryl; (hydroxy)aryl,((carboxy)alkyl)aryl; (((dialkyl)amino)alkyl)aryl; (nitroalkyl)aryl;(((alkylsulfonyl)amino)alkyl)aryl; ((heterocycloaliphatic)carbonyl)aryl;((alkylsulfonyl)alkyl)aryl; (cyanoalkyl)aryl; (hydroxyalkyl)aryl;(alkylcarbonyl)aryl; alkylaryl; (trihaloalkyl)aryl;p-amino-m-alkoxycarbonylaryl; p-amino-m-cyanoaryl; p-halo-m-aminoaryl;or (m-(heterocycloaliphatic)-o-(alkyl))aryl.

As used herein, an “araliphatic” such as an “aralkyl” group refers to analiphatic group (e.g., a C₁₋₄ alkyl group) that is substituted with anaryl group. “Aliphatic”, “alkyl”, and “aryl” are defined herein. Anexample of an araliphatic such as an aralkyl group is benzyl.

As used herein, an “aralkyl” group refers to an alkyl group (e.g., aC₁₋₄ alkyl group) that is substituted with an aryl group. Both “alkyl”and “aryl” have been defined above. An example of an aralkyl group isbenzyl. An aralkyl is optionally substituted with one or moresubstituents such as aliphatic [e.g., alkyl, alkenyl, or alkynyl,including carboxyalkyl, hydroxyalkyl, or haloalkyl such astrifluoromethyl], cycloaliphatic [e.g., cycloalkyl or cycloalkenyl],(cycloalkyl)alkyl, heterocycloalkyl, (heterocycloalkyl)alkyl, aryl,heteroaryl, alkoxy, cycloalkyloxy, heterocycloalkyloxy, aryloxy,heteroaryloxy, aralkyloxy, heteroaralkyloxy, aroyl, heteroaroyl, nitro,carboxy, alkoxycarbonyl, alkylcarbonyloxy, amido [e.g., aminocarbonyl,alkylcarbonylamino, cycloalkylcarbonylamino,(cycloalkylalkyl)carbonylamino, arylcarbonylamino, aralkylcarbonylamino,(heterocycloalkyl)carbonylamino, (heterocycloalkylalkyl)carbonylamino,heteroarylcarbonylamino, or heteroaralkylcarbonylamino], cyano, halo,hydroxy, acyl, mercapto, alkylsulfanyl, sulfoxy, urea, thiourea,sulfamoyl, sulfamide, oxo, or carbamoyl.

As used herein, a “bicyclic ring system” includes 6-12 (e.g., 8-12 or 9,10, or 11) membered structures that form two rings, wherein the tworings have at least one atom in common (e.g., 2 atoms in common).Bicyclic ring systems include bicycloaliphatics (e.g., bicycloalkyl orbicycloalkenyl), bicycloheteroaliphatics, bicyclic aryls, and bicyclicheteroaryls.

As used herein, a “cycloaliphatic” group encompasses a “cycloalkyl”group and a “cycloalkenyl” group, each of which being optionallysubstituted as set forth below.

As used herein, a “cycloalkyl” group refers to a saturated carbocyclicmono- or bicyclic (fused or bridged) ring of 3-10 (e.g., 5-10) carbonatoms. Examples of cycloalkyl groups include cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, cycloheptyl, adamantyl, norbornyl, cubyl,octahydro-indenyl, decahydro-naphthyl, bicyclo[3.2.1]octyl,bicyclo[2.2.2]octyl, bicyclo[3.3.1]nonyl, bicyclo[3.3.2.]decyl,bicyclo[2.2.2]octyl, adamantyl, or((aminocarbonyl)cycloalkyl)cycloalkyl.

A “cycloalkenyl” group, as used herein, refers to a non-aromaticcarbocyclic ring of 3-10 (e.g., 4-8) carbon atoms having one or moredouble bonds. Examples of cycloalkenyl groups include cyclopentenyl,1,4-cyclohexa-di-enyl, cycloheptenyl, cyclooctenyl, hexahydro-indenyl,octahydro-naphthyl, cyclohexenyl, bicyclo[2.2.2]octenyl, orbicyclo[3.3.1]nonenyl.

A cycloalkyl or cycloalkenyl group can be optionally substituted withone or more substituents such as phospho, aliphatic [e.g., alkyl,alkenyl, or alkynyl], cycloaliphatic, (cycloaliphatic) aliphatic,heterocycloaliphatic, (heterocycloaliphatic) aliphatic, aryl,heteroaryl, alkoxy, (cycloaliphatic)oxy, (heterocycloaliphatic)oxy,aryloxy, heteroaryloxy, (araliphatic)oxy, (heteroaraliphatic)oxy, aroyl,heteroaroyl, amino, amido [e.g., (aliphatic)carbonylamino,(cycloaliphatic)carbonylamino, ((cycloaliphatic)aliphatic)carbonylamino,(aryl)carbonylamino, (araliphatic)carbonylamino,(heterocycloaliphatic)carbonylamino,((heterocycloaliphatic)aliphatic)carbonylamino,(heteroaryl)carbonylamino, or (heteroaraliphatic)carbonylamino], nitro,carboxy [e.g., HOOC—, alkoxycarbonyl, or alkylcarbonyloxy], acyl [e.g.,(cycloaliphatic)carbonyl, ((cycloaliphatic) aliphatic)carbonyl,(araliphatic)carbonyl, (heterocycloaliphatic)carbonyl,((heterocycloaliphatic)aliphatic)carbonyl, or(heteroaraliphatic)carbonyl], cyano, halo, hydroxy, mercapto, sulfonyl[e.g., alkyl-SO₂— and aryl-SO₂—], sulfinyl [e.g., alkyl-S(O)—], sulfanyl[e.g., alkyl-S—], sulfoxy, urea, thiourea, sulfamoyl, sulfamide, oxo, orcarbamoyl.

As used herein, the term “heterocycloaliphatic” encompassesheterocycloalkyl groups and heterocycloalkenyl groups, each of whichbeing optionally substituted as set forth below.

As used herein, a “heterocycloalkyl” group refers to a 3-10 memberedmono- or bicylic (fused or bridged) (e.g., 5- to 10-membered mono- orbicyclic) saturated ring structure, in which one or more of the ringatoms is a heteroatom (e.g., N, O, S, or combinations thereof). Examplesof a heterocycloalkyl group include piperidyl, piperazyl,tetrahydropyranyl, tetrahydrofuryl, 1,4-dioxolanyl, 1,4-dithianyl,1,3-dioxolanyl, oxazolidyl, isoxazolidyl, morpholinyl, thiomorpholyl,octahydrobenzofuryl, octahydrochromenyl, octahydrothiochromenyl,octahydroindolyl, octahydropyrindinyl, decahydroquinolinyl,octahydrobenzo[b]thiopheneyl, 2-oxa-bicyclo[2.2.2]octyl,1-aza-bicyclo[2.2.2]octyl, 3-aza-bicyclo[3.2.1]octyl, and2,6-dioxa-tricyclo[3.3.1.0^(3,7)]nonyl. A monocyclic heterocycloalkylgroup can be fused with a phenyl moiety to form structures, such astetrahydroisoquinoline, which would be categorized as heteroaryls.

A “heterocycloalkenyl” group, as used herein, refers to a mono- orbicylic (e.g., 5- to 10-membered mono- or bicyclic) non-aromatic ringstructure having one or more double bonds, and wherein one or more ofthe ring atoms is a heteroatom (e.g., N, O, or S). Monocyclic andbicyclic heterocycloaliphatics are numbered according to standardchemical nomenclature.

A heterocycloalkyl or heterocycloalkenyl group can be optionallysubstituted with one or more substituents such as phospho, aliphatic[e.g., alkyl, alkenyl, or alkynyl], cycloaliphatic,(cycloaliphatic)aliphatic, heterocycloaliphatic,(heterocycloaliphatic)aliphatic, aryl, heteroaryl, alkoxy,(cycloaliphatic)oxy, (heterocycloaliphatic)oxy, aryloxy, heteroaryloxy,(araliphatic)oxy, (heteroaraliphatic)oxy, aroyl, heteroaroyl, amino,amido [e.g., (aliphatic)carbonylamino, (cycloaliphatic)carbonylamino,((cycloaliphatic) aliphatic)carbonylamino, (aryl)carbonylamino,(araliphatic)carbonylamino, (heterocycloaliphatic)carbonylamino,((heterocycloaliphatic) aliphatic)carbonylamino,(heteroaryl)carbonylamino, or (heteroaraliphatic)carbonylamino], nitro,carboxy [e.g., HOOC—, alkoxycarbonyl, or alkylcarbonyloxy], acyl [e.g.,(cycloaliphatic)carbonyl, ((cycloaliphatic) aliphatic)carbonyl,(araliphatic)carbonyl, (heterocycloaliphatic)carbonyl,((heterocycloaliphatic)aliphatic)carbonyl, or(heteroaraliphatic)carbonyl], nitro, cyano, halo, hydroxy, mercapto,sulfonyl [e.g., alkylsulfonyl or arylsulfonyl], sulfinyl [e.g.,alkylsulfinyl], sulfanyl [e.g., alkylsulfanyl], sulfoxy, urea, thiourea,sulfamoyl, sulfamide, oxo, or carbamoyl.

A “heteroaryl” group, as used herein, refers to a monocyclic, bicyclic,or tricyclic ring system having 4 to 15 ring atoms wherein one or moreof the ring atoms is a heteroatom (e.g., N, O, S, or combinationsthereof) and in which the monocyclic ring system is aromatic or at leastone of the rings in the bicyclic or tricyclic ring systems is aromatic.A heteroaryl group includes a benzofused ring system having 2 to 3rings. For example, a benzofused group includes benzo fused with one ortwo 4 to 8 membered heterocycloaliphatic moieties (e.g., indolizyl,indolyl, isoindolyl, 3H-indolyl, indolinyl, benzo[b]furyl,benzo[b]thiophene-yl, quinolinyl, or isoquinolinyl). Some examples ofheteroaryl are azetidinyl, pyridyl, 1H-indazolyl, furyl, pyrrolyl,thienyl, thiazolyl, oxazolyl, imidazolyl, tetrazolyl, benzofuryl,isoquinolinyl, benzthiazolyl, xanthene, thioxanthene, phenothiazine,dihydroindole, benzo[1,3]dioxole, benzo[b]furyl, benzo[b]thiophenyl,indazolyl, benzimidazolyl, benzthiazolyl, puryl, cinnolyl, quinolyl,quinazolyl, cinnolyl, phthalazyl, quinazolyl, quinoxalyl, isoquinolyl,4H-quinolizyl, benzo-1,2,5-thiadiazolyl, or 1,8-naphthyridyl.

Without limitation, monocyclic heteroaryls include furyl, thiophene-yl,2H-pyrrolyl, pyrrolyl, oxazolyl, thazolyl, imidazolyl, pyrazolyl,isoxazolyl, isothiazolyl, 1,3,4-thiadiazolyl, 2H-pyranyl, 4-H-pranyl,pyridyl, pyridazyl, pyrimidyl, pyrazolyl, pyrazyl, or 1,3,5-triazyl.Monocyclic heteroaryls are numbered according to standard chemicalnomenclature.

Without limitation, bicyclic heteroaryls include indolizyl, indolyl,isoindolyl, 3H-indolyl, indolinyl, benzo[b]furyl, benzo[b]thiophenyl,quinolinyl, isoquinolinyl, indolizyl, isoindolyl, indolyl,benzo[b]furyl, bexo[b]thiophenyl, indazolyl, benzimidazyl,benzthiazolyl, purinyl, 4H-quinolizyl, quinolyl, isoquinolyl, cinnolyl,phthalazyl, quinazolyl, quinoxalyl, 1,8-naphthyridyl, or pteridyl.Bicyclic heteroaryls are numbered according to standard chemicalnomenclature.

A heteroaryl is optionally substituted with one or more substituentssuch as aliphatic [e.g., alkyl, alkenyl, or alkynyl]; cycloaliphatic;(cycloaliphatic)aliphatic; heterocycloaliphatic;(heterocycloaliphatic)aliphatic; aryl; heteroaryl; alkoxy;(cycloaliphatic)oxy; (heterocycloaliphatic)oxy; aryloxy; heteroaryloxy;(araliphatic)oxy; (heteroaraliphatic)oxy; aroyl; heteroaroyl; amino; oxo(on a non-aromatic carbocyclic or heterocyclic ring of a bicyclic ortricyclic heteroaryl); carboxy; amido; acyl [e.g., aliphaticcarbonyl;(cycloaliphatic)carbonyl; ((cycloaliphatic)aliphatic)carbonyl;(araliphatic)carbonyl; (heterocycloaliphatic)carbonyl;((heterocycloaliphatic)aliphatic)carbonyl; or(heteroaraliphatic)carbonyl]; sulfonyl [e.g., aliphaticsulfonyl oraminosulfonyl]; sulfinyl [e.g., aliphaticsulfinyl]; sulfanyl [e.g.,aliphaticsulfanyl]; nitro; cyano; halo; hydroxy; mercapto; sulfoxy;urea; thiourea; sulfamoyl; sulfamide; or carbamoyl. Alternatively, aheteroaryl can be unsubstituted.

Non-limiting examples of substituted heteroaryls include(halo)heteroaryl [e.g., mono- and di-(halo)heteroaryl];(carboxy)heteroaryl [e.g., (alkoxycarbonyl)heteroaryl]; cyanoheteroaryl;aminoheteroaryl [e.g., ((alkylsulfonyl)amino)heteroaryl and((dialkyl)amino)heteroaryl]; (amido)heteroaryl [e.g.,aminocarbonylheteroaryl, ((alkylcarbonyl)amino)heteroaryl,((((alkyl)amino)alkyl)aminocarbonyl)heteroaryl,(((heteroaryl)amino)carbonyl)heteroaryl,((heterocycloaliphatic)carbonyl)heteroaryl, and((alkylcarbonyl)amino)heteroaryl]; (cyanoalkyl)heteroaryl;(alkoxy)heteroaryl; (sulfamoyl)heteroaryl [e.g.,(aminosulfonyl)heteroaryl]; (sulfonyl)heteroaryl [e.g.,(alkylsulfonyl)heteroaryl]; (hydroxyalkyl)heteroaryl;(alkoxyalkyl)heteroaryl; (hydroxy)heteroaryl;((carboxy)alkyl)heteroaryl; (((dialkyl)amino)alkyl]heteroaryl;(heterocycloaliphatic)heteroaryl; (cycloaliphatic)heteroaryl;(nitroalkyl)heteroaryl; (((alkylsulfonyl)amino)alkyl)heteroaryl;((alkylsulfonyl)alkyl)heteroaryl; (cyanoalkyl)heteroaryl;(acyl)heteroaryl [e.g., (alkylcarbonyl)heteroaryl]; (alkyl)heteroaryl;or (haloalkyl)heteroaryl [e.g., trihaloalkylheteroaryl].

A “heteroaraliphatic (such as a heteroaralkyl group) as used herein,refers to an aliphatic group (e.g., a C₁₋₄ alkyl group) that issubstituted with a heteroaryl group. “Aliphatic”, “alkyl”, and“heteroaryl” have been defined above.

A “heteroaralkyl” group, as used herein, refers to an alkyl group (e.g.,a C₁₋₄ alkyl group) that is substituted with a heteroaryl group. Both“alkyl” and “heteroaryl” have been defined above. A heteroaralkyl isoptionally substituted with one or more substituents such as alkyl(including carboxyalkyl, hydroxyalkyl, and haloalkyl such astrifluoromethyl), alkenyl, alkynyl, cycloalkyl, (cycloalkyl)alkyl,heterocycloalkyl, (heterocycloalkyl)alkyl, aryl, heteroaryl, alkoxy,cycloalkyloxy, heterocycloalkyloxy, aryloxy, heteroaryloxy, aralkyloxy,heteroaralkyloxy, aroyl, heteroaroyl, nitro, carboxy, alkoxycarbonyl,alkylcarbonyloxy, aminocarbonyl, alkylcarbonylamino,cycloalkylcarbonylamino, (cycloalkylalkyl)carbonylamino,arylcarbonylamino, aralkylcarbonylamino,(heterocycloalkyl)carbonylamino, (heterocycloalkylalkyl)carbonylamino,heteroarylcarbonylamino, heteroaralkylcarbonylamino, cyano, halo,hydroxy, acyl, mercapto, alkylsulfanyl, sulfoxy, urea, thiourea,sulfamoyl, sulfamide, oxo, or carbamoyl.

As used herein, “cyclic moiety” and “cyclic group” refer to mono-, bi-,and tri-cyclic ring systems including cycloaliphatic,heterocycloaliphatic, aryl, or heteroaryl, each of which has beenpreviously defined.

As used herein, a “bridged bicyclic ring system” refers to a bicyclicheterocyclicalipahtic ring system or bicyclic cycloaliphatic ring systemin which the rings are bridged. Examples of bridged bicyclic ringsystems include, but are not limited to, adamantanyl, norbornanyl,bicyclo[3.2.1]octyl, bicyclo[2.2.2]octyl, bicyclo[3.3.1]nonyl,bicyclo[3.3.2]decyl, 2-oxabicyclo[2.2.2]octyl, 1-azabicyclo[2.2.2]octyl,3-azabicyclo[3.2.1]octyl, and 2,6-dioxa-tricyclo[3.3.1.0^(3,7)]nonyl. Abridged bicyclic ring system can be optionally substituted with one ormore substituents such as alkyl (including carboxyalkyl, hydroxyalkyl,and haloalkyl such as trifluoromethyl), alkenyl, alkynyl, cycloalkyl,(cycloalkyl)alkyl, heterocycloalkyl, (heterocycloalkyl)alkyl, aryl,heteroaryl, alkoxy, cycloalkyloxy, heterocycloalkyloxy, aryloxy,heteroaryloxy, aralkyloxy, heteroaralkyloxy, aroyl, heteroaroyl, nitro,carboxy, alkoxycarbonyl, alkylcarbonyloxy, aminocarbonyl,alkylcarbonylamino, cycloalkylcarbonylamino,(cycloalkylalkyl)carbonylamino, arylcarbonylamino, aralkylcarbonylamino,(heterocycloalkyl)carbonylamino, (heterocycloalkylalkyl)carbonylamino,heteroarylcarbonylamino, heteroaralkylcarbonylamino, cyano, halo,hydroxy, acyl, mercapto, alkylsulfanyl, sulfoxy, urea, thiourea,sulfamoyl, sulfamide, oxo, or carbamoyl.

As used herein, an “acyl” group refers to a formyl group or R^(X)—C(O)—(such as alkyl-C(O)—, also referred to as “alkylcarbonyl”) where R^(X)and “alkyl” have been defined previously. Acetyl and pivaloyl areexamples of acyl groups.

As used herein, an “aroyl” or “heteroaroyl” refers to an aryl-C(O)— or aheteroaryl-C(O)—. The aryl and heteroaryl portion of the aroyl orheteroaroyl is optionally substituted as previously defined.

As used herein, an “alkoxy” group refers to an alkyl-O— group where“alkyl” has been defined previously.

As used herein, a “carbamoyl” group refers to a group having thestructure —O—CO—NR^(X)R^(Y) or —NR^(X)—CO—O—R^(Z), wherein R^(X) andR^(Y) have been defined above and R^(Z) can be aliphatic, aryl,araliphatic, heterocycloaliphatic, heteroaryl, or heteroaraliphatic.

As used herein, a “carboxy” group refers to —COOH, —COOR^(X), —OC(O)H,—OC(O)R^(X), when used as a terminal group; or —OC(O)— or —C(O)O— whenused as an internal group.

As used herein, a “haloaliphatic” group refers to an aliphatic groupsubstituted with 1-3 halogen. For instance, the term haloalkyl includesthe group —CF₃.

As used herein, a “mercapto” group refers to —SH.

As used herein, a “sulfo” group refers to —SO₃H or —SO₃R^(X) when usedterminally or —S(O)₃— when used internally.

As used herein, a “sulfamide” group refers to the structure—NR^(X)—S(O)₂—NR^(Y)R^(Z) when used terminally and —NR^(X)—S(O)₂—NR^(Y)—when used internally, wherein R^(X), R^(Y), and R^(Z) have been definedabove.

As used herein, a “sulfamoyl” group refers to the structure—O—S(O)₂—NR^(Y)R^(Z) wherein R^(Y) and R^(Z) have been defined above.

As used herein, a “sulfonamide” group refers to the structure—S(O)₂—NR^(X)R^(Y) or —NR^(X)—S(O)₂—R^(Z) when used terminally; or—S(O)₂—NR^(X)— or —NR^(X)—S(O)₂— when used internally, wherein R^(X),R^(Y), and R^(Z) are defined above.

As used herein a “sulfanyl” group refers to —S—R^(X) when usedterminally and —S— when used internally, wherein R^(X) has been definedabove. Examples of sulfanyls include aliphatic-S—, cycloaliphatic-S—,aryl-S—, or the like.

As used herein a “sulfinyl” group refers to —S(O)—R^(X) when usedterminally and —S(O)—when used internally, wherein R^(X) has beendefined above. Exemplary sulfinyl groups include aliphatic-S(O)—,aryl-S(O)—, (cycloaliphatic(aliphatic))-S(O)—, cycloalkyl-S(O)—,heterocycloaliphatic-S(O)—, heteroaryl-S(O)—, or the like.

As used herein, a “sulfonyl” group refers to —S(O)₂—R^(X) when usedterminally and —S(O)₂— when used internally, wherein R^(X) has beendefined above. Exemplary sulfonyl groups include aliphatic-S(O)₂—,aryl-S(O)₂—, (cycloaliphatic(aliphatic))-S(O)₂—, cycloaliphatic-S(O)₂—,heterocycloaliphatic-S(O)₂—, heteroaryl-S(O)₂—,(cycloaliphatic(amido(aliphatic)))-S(O)₂— or the like.

As used herein, a “sulfoxy” group refers to —O—S(O)—R^(X) or—S(O)—O—R^(X), when used terminally and —O—S(O)— or —S(O)—O— when usedinternally, where R^(X) has been defined above.

As used herein, a “halogen” or “halo” group refers to fluorine,chlorine, bromine or iodine.

As used herein, an “alkoxycarbonyl”, which is encompassed by the termcarboxy, used alone or in connection with another group refers to agroup such as alkyl-O—C(O)—.

As used herein, an “alkoxyalkyl” refers to an alkyl group such asalkyl-O-alkyl-, wherein alkyl has been defined above.

As used herein, a “carbonyl” refer to —C(O)—.

As used herein, an “oxo” refers to ═O.

As used herein, the term “phospho” refers to phosphinates andphosphonates. Examples of phosphinates and phosphonates include—P(O)(R^(P))₂, wherein R^(P) is aliphatic, alkoxy, aryloxy,heteroaryloxy, (cycloaliphatic)oxy, (heterocycloaliphatic)oxy aryl,heteroaryl, cycloaliphatic or amino.

As used herein, an “aminoalkyl” refers to the structure(R^(X))₂N-alkyl-.

As used herein, a “cyanoalkyl” refers to the structure (NC)-alkyl-.

As used herein, a “urea” group refers to the structure—NR^(X)—CO—NR^(Y)R^(Z) and a “thiourea” group refers to the structure—NR^(X)—CS—NR^(Y)R^(Z) when used terminally and —NR^(X)—CO—NR^(Y)— or—NR^(X)—CS—NR^(Y)— when used internally, wherein R^(X), R^(Y), and R^(Z)have been defined above.

As used herein, a “guanidine” group refers to the structure—N═C(N(R^(X)R^(Y)))N(R^(X)R^(Y)) or —NR^(X)—C(═NR^(X))NR^(X)R^(Y)wherein R^(X) and R^(Y) have been defined above.

As used herein, the term “amidino” group refers to the structure—C═(NR^(X))N(R^(X)R^(Y)) wherein R^(X) and R^(Y) have been definedabove.

In general, the term “vicinal” refers to the placement of substituentson a group that includes two or more carbon atoms, wherein thesubstituents are attached to adjacent carbon atoms.

In general, the term “geminal” refers to the placement of substituentson a group that includes two or more carbon atoms, wherein thesubstituents are attached to the same carbon atom.

The terms “terminally” and “internally” refer to the location of a groupwithin a substituent. A group is terminal when the group is present atthe end of the substituent not further bonded to the rest of thechemical structure. Carboxyalkyl, i.e., R^(X)O(O)C-alkyl is an exampleof a carboxy group used terminally. A group is internal when the groupis present in the middle of a substituent of the chemical structure.Alkylcarboxy (e.g., alkyl-C(O)O— or alkyl-OC(O)—) and alkylcarboxyaryl(e.g., alkyl-C(O)O-aryl- or alkyl-O(CO)-aryl-) are examples of carboxygroups used internally.

As used herein, “relative retention time” refers to the ratio betweenthe net retention time of a substance and that of a standard compound.

As used herein, an “aliphatic chain” refers to a branched or straightaliphatic group (e.g., alkyl groups, alkenyl groups, or alkynyl groups).A straight aliphatic chain has the structure —[CH₂]_(v)—, where v is1-12. A branched aliphatic chain is a straight aliphatic chain that issubstituted with one or more aliphatic groups. A branched aliphaticchain has the structure —[CQQ]_(v)- where Q is independently a hydrogenor an aliphatic group; however, Q shall be an aliphatic group in atleast one instance. The term aliphatic chain includes alkyl chains,alkenyl chains, and alkynyl chains, where alkyl, alkenyl, and alkynylare defined above.

In general, the term “substituted”, whether preceded by the term“optionally” or not, refers to the replacement of hydrogen atoms in agiven structure with the radical of a specified substituent. Specificsubstituents are described above in the definitions and below in thedescription of compounds and examples thereof. Unless otherwiseindicated, an optionally substituted group can have a substituent ateach substitutable position of the group, and when more than oneposition in any given structure can be substituted with more than onesubstituent selected from a specified group, the substituent can beeither the same or different at every position. A ring substituent, suchas a heterocycloalkyl, can be bound to another ring, such as acycloalkyl, to form a spiro-bicyclic ring system, e.g., both rings shareone common atom. As one of ordinary skill in the art will recognize,combinations of substituents envisioned by this invention are thosecombinations that result in the formation of stable or chemicallyfeasible compounds.

The phrase “stable or chemically feasible”, as used herein, refers tocompounds that are not substantially altered when subjected toconditions to allow for their production, detection, and preferablytheir recovery, purification, and use for one or more of the purposesdisclosed herein. In some embodiments, a stable compound or chemicallyfeasible compound is one that is not substantially altered when kept ata temperature of 40° C. or less, in the absence of moisture or otherchemically reactive conditions, for at least a week.

As used herein, an “effective amount” is defined as the amount requiredto confer a therapeutic effect on the treated patient, and is typicallydetermined based on age, surface area, weight, and condition of thepatient. The interrelationship of dosages for animals and humans (basedon milligrams per meter squared of body surface) is described byFreireich et al., Cancer Chemother. Rep., 50: 219 (1966). Body surfacearea may be approximately determined from height and weight of thepatient. See, e.g., Scientific Tables, Geigy Pharmaceuticals, Ardsley,N.Y., 537 (1970). As used herein, “patient” refers to a mammal,including a human.

Chemical structures and nomenclature are derived from ChemDraw, version11.0.1, Cambridge, Mass.

It is noted that the use of the descriptors “first”, “second”, “third”,or the like is used to differentiate separate elements (e.g., solvents,reaction steps, processes, reagents, or the like) and may or may notrefer to the relative order or relative chronology of the elementsdescribed.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts which are, within the scope of sound medical judgement,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like.

Pharmaceutically acceptable salts are well known in the art. Forexample, S. M. Berge et al., describe pharmaceutically acceptable saltsin detail in J. Pharmaceutical Sciences, 1977, 66, 1-19, incorporatedherein by reference. Pharmaceutically acceptable salts of the compoundsof this invention include those derived from suitable inorganic andorganic acids and bases. Examples of pharmaceutically acceptable,nontoxic acid addition salts are salts of an amino group formed withinorganic acids such as hydrochloric acid, hydrobromic acid, phosphoricacid, sulfuric acid and perchloric acid or with organic acids such asacetic acid, oxalic acid, maleic acid, tartaric acid, citric acid,succinic acid or malonic acid or by using other methods used in the artsuch as ion exchange. Other pharmaceutically acceptable salts includeadipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate,bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate,pivalate, propionate, stearate, succinate, sulfate, tartrate,thiocyanate, p-toluenesulfonate, undecanoate, valerate salts, and thelike. Salts derived from appropriate bases include alkali metal,alkaline earth metal, ammonium and N⁺(C₁₋₄alkyl)₄ salts. This inventionalso envisions the quaternization of any basic nitrogen-containinggroups of the compounds disclosed herein. Water or oil-soluble ordispersible products may be obtained by such quaternization.Representative alkali or alkaline earth metal salts include sodium,lithium, potassium, calcium, magnesium, and the like. Furtherpharmaceutically acceptable salts include, when appropriate, nontoxicammonium, quaternary ammonium, and amine cations formed usingcounterions such as halide, hydroxide, carboxylate, sulfate, phosphate,nitrate, loweralkyl sulfonate and aryl sulfonate.

As described herein, “protecting group” refers to a moiety orfunctionality that is introduced into a molecule by chemicalmodification of a functional group in order to obtain chemoselectivityin a subsequent chemical reaction. Standard protecting groups areprovided in Wuts and Greene: “Greene's Protective Groups in OrganicSynthesis” 4th Ed, Wuts, P. G. M. and Greene, T. W., Wiley-Interscience,New York: 2006, which is incorporated herein by reference.

Examples of nitrogen protecting groups include acyl, aroyl, or carbamylgroups such as formyl, acetyl, propionyl, pivaloyl, t-butylacetyl,2-chloroacetyl, 2-bromoacetyl, trifluoroacetyl, trichloroacetyl,phthalyl, o-nitrophenoxyacetyl, α-chlorobutyryl, benzoyl,4-chlorobenzoyl, 4-bromobenzoyl, 4-nitrobenzoyl and chiral auxiliariessuch as protected or unprotected D, L or D, L-amino acids such asalanine, leucine, phenylalanine and the like; sulfonyl groups such asbenzenesulfonyl, p-toluenesulfonyl and the like; carbamate groups suchas benzyloxycarbonyl, p-chlorobenzyloxycarbonyl,p-methoxybenzyloxycarbonyl, p-nitrobenzyloxycarbonyl,2-nitrobenzyloxycarbonyl, p-bromobenzyloxycarbonyl,3,4-dimethoxybenzyloxycarbonyl, 3,5-dimethoxybenzyloxycarbonyl,2,4-dimethoxybenzyloxycarbonyl, 4-methoxybenzyloxycarbonyl,2-nitro-4,5-dimethoxybenzyloxycarbonyl,3,4,5-trimethoxybenzyloxycarbonyl,1-(p-biphenylyl)-1-methylethoxycarbonyl,α,α-dimethyl-3,5-dimethoxybenzyloxycarbonyl, benzhydryloxycarbonyl,t-butyloxycarbonyl, diisopropylmethoxycarbonyl, isopropyloxycarbonyl,ethoxycarbonyl, methoxycarbonyl, allyloxycarbonyl,2,2,2,-trichloroethoxycarbonyl, phenoxycarbonyl, 4-nitrophenoxycarbonyl, fluorenyl-9-methoxycarbonyl, cyclopentyloxycarbonyl,adamantyloxycarbonyl, cyclohexyloxycarbonyl, phenylthiocarbonyl and thelike, arylalkyl groups such as benzyl, triphenylmethyl, benzyloxymethyland the like and silyl groups such as trimethylsilyl and the like.Preferred N-protecting groups are benzenesulfonylchloridep-toluenesulfonyl and the like, including, but not limited to, tosyl.

Unless otherwise stated, structures depicted herein are also meant toinclude all isomeric (e.g., enantiomeric, diastereomeric, and geometric(or conformational)) forms of the structure; for example, the R and Sconfigurations for each asymmetric center, (Z) and (E) double bondisomers, and (Z) and (E) conformational isomers. Therefore, singlestereochemical isomers as well as enantiomeric, diastereomeric, andgeometric (or conformational) mixtures of the present compounds arewithin the scope of the invention. Unless otherwise stated, alltautomeric forms of the compounds of the invention are within the scopeof the invention. Additionally, unless otherwise stated, structuresdepicted herein are also meant to include compounds that differ only inthe presence of one or more isotopically enriched atoms. For example,compounds having the present structures except for the replacement ofhydrogen by deuterium or tritium, or the replacement of a carbon by a¹³C- or ¹⁴C-enriched carbon are within the scope of this invention. Suchcompounds are useful, for example, as analytical tools, probes inbiological assays, or JAK inhibitors with improved therapeutic profile.

As used herein, the term “solvent” also includes mixtures of solvents.

III. PREPARATION OF COMPOUNDS

It is noted that the steps recited herein may be performed in anychronological order without regard to step letter or number. Forexample, step v) may precede or follow step (g), step (e), step (f), orstep (s). Likewise, step i) may precede or follow step ii), step iii),step iv), or step v).

The compound of Formula (I) has the following structure:

wherein R¹ is —Cl or —F. Compounds of Formula (I) and pharmaceuticallyacceptable salts thereof are inhibitors of the replication of influenzaviruses, and can be used for treating influenza in a patient, asdescribed in WO 2013/019828. In one specific embodiment, R^(L) is —F. Inanother specific embodiment, R¹ is —Cl.

In one embodiment, the invention is directed to a process for preparinga compound of Formula I:

or a pharmaceutically acceptable salt thereof, wherein R¹ is —Cl or —F;comprising the step of:

i) reacting a compound of Formula 1 with a compound of Formula 2

in the presence of water, an organic solvent, a base, and a transitionmetal catalyst to generate a compound of Formula I; wherein

each of R⁸ and R⁹ is —OH, —O—C₁₋₄ aliphatic optionally substituted with1-5 occurrences of R¹¹, or

-   -   R⁸ and R⁹ together with the boron atom to which they are        attached, form a 5-9 membered mono- or bicyclic ring system,        optionally substituted with 1-6 occurrences of R¹¹, or BF₃K;    -   each R¹¹ is independently selected from halogen, —OCH₃, —OH,        —NO₂, —NH₂, —SH, —SCH₃, —NHCH₃, —CN, ═O, or unsubstituted —C₁₋₂        aliphatic;    -   X¹ is —Cl, —Br, —I, —OTs, —OMs, —OH, or —NH₂; and    -   the transition metal catalyst comprises Pd.

In some embodiments, the base of step i) is an organic base. Forexample, the organic base comprises a tertiary amine. And, in someinstances, the tertiary amine comprises diisopropylethylamine,triethylamine, triethylenediamine, or any combination thereof.

In some embodiments, the organic solvent of step i) is an aproticsolvent. For example, the aprotic solvent is acetonitrile, toluene,N,N-dimethylformamide, N,N-dimethylacetamide, acetone, methyl tert-butylether, or any combination thereof.

In some embodiments, R¹ is —F. In other embodiments, R¹ is —Cl.

In some embodiments, X¹ is —Cl.

In some embodiments, the transition metal catalyst comprisespalladium(II)acetate, tetrakis(triphenylphosphine)palladium(0),tris(dibenzylideneacetone)dipalladium(0), or any combination thereof. Insome embodiments, the transition metal catalyst is palladium(II)acetate.Other examples of transition metal catalysts comprising Pd include

or, or any combination thereof.

In some embodiments, the transition metal catalyst is

or the transition metal catalyst comprises Pd and a ligand comprising

In some embodiments, the palladium catalyst is formed in situ.

In some embodiments, the reaction of step i) is performed at atemperature between about 50° C. and about 110° C. For example, thereaction of step i) is performed at a temperature between about 60° C.and about 95° C. In other examples, the reaction of step i) is performedat a temperature between about 65° C. and about 80° C.

Another embodiment of the present invention provides a method ofpreparing a compound of Formula I

or a pharmaceutically acceptable salt thereof wherein R¹ is —Cl or —F;comprising comprising the step of:

x) reacting a compound of Formula 1 with a compound of Formula 2

in the presence of water, an organic solvent, a base, and a transitionmetal catalyst to generate a compound of Formula 3;

wherein

each of R⁸ and R⁹ is —OH, —O—C₁₋₄ aliphatic optionally substituted with1-5 occurrences of R^(1′), or

-   -   R⁸ and R⁹ together with the boron atom to which they are        attached, form a 5-9 membered mono- or bicyclic ring system,        optionally substituted with 1-6 occurrences of R¹¹, or BF₃K;

each R¹¹ is independently selected from halogen, —OCH₃, —OH, —NO₂, —NH₂,—SH, —SCH₃, —NHCH₃, —CN, ═O, or unsubstituted —C₁₋₂ aliphatic;

X¹ is —Cl, —Br, —I, —OTs, —OMs, —OH, or —NH₂; and

the transition metal catalyst is

or the transition metal catalyst comprises Pd and a ligand comprising

and

ii) deprotecting the compound of Formula 3 to form a compound of FormulaI.

In some embodiments, the palladium catalyst of either of steps i) or x)is formed in situ. And, in other embodiments, the palladium-AmPhoscomplex is employed as a pre-prepared catalyst. Typical examples ofPd(0) or Pd(II) sources include Pd₂(dba)₃, Pd(OAc)₂, and PdCl₂, whereindba is dibenzyllideneacetone and OAc is acetate. In one specificembodiment, the palladium-AmPhos complex is prepared in situ by mixingPdCl₂ and AmPhos.

In some embodiments, the reaction of either of steps i) or x) isperformed at a temperature between about 50° C. and about 110° C. Forexample, the reaction of either of steps i) or x) is performed at atemperature between about 60° C. and about 95° C. In other examples, thereaction of either of steps i) or x) is performed at a temperaturebetween about 70° C. and about 80° C.

In some embodiments, the base of either of steps i) or x) is an organicbase. For example, the organic base comprises a tertiary amine. And, insome instances, the tertiary amine comprises diisopropylethylamine,triethylamine, triethylenediamine, or any combination thereof. Alkalimetal hydroxides (e.g., NaOH and/or KOH), alkali metal carbonates (e.g.K₂CO₃, and/or Na₂CO₃), or alkali metal alkoxides (e.g., NaO^(t)Bu and/orKO^(t)Bu) may also be used as the base for the reaction of either ofsteps i) or x).

The reaction of either of steps i) or x) can be performed in thepresence of any suitable solvent system. Examples of suitable solventsystems include aqueous solvent systems. In some embodiments, theorganic solvent of either of steps i) or x) is an aprotic solvent. Forexample, the aprotic solvent is acetonitrile, toluene,N,N-dimethylformamide (DMF), N,N-dimethylacetamide (DMA), acetone,methyl tert-butyl ether, or any combination thereof. In another example,the solvent comprises water and at least one organic solvent selectedfrom DMF, IPA, toluene, acetonitrile, THF, DME, and dioxane.

Some embodiments further include one or more recrystallization steps.Some instances further include step xi) reacting the compound of Formula3 with p-toluenesulfonic acid (TsOH) or other acid in the presence of asolvent system to generate the p-toluenesulfonic acid (TsOH) salt of thecompound of Formula 3; and step xii) treating the p-toluenesulfonic acid(TsOH) salt of the compound of Formula 3 with an inorganic base togenerate the compound of Formula I.

The deprotection step ii) can be performed in any suitable conditionsknown in the art for deprotection of a tosyl protecting group. In onespecific embodiment, the deprotection step employs treating the compoundof Formula 3 or a pharmaceutically acceptable salt thereof with aninorganic hydroxide. Typical examples of suitable inorganic hydroxidesinclude LiOH, NaOH, and KOH. In one specific embodiment, LiOH isemployed. In another specific embodiment, the deprotection step ii)employs LiOH in a solvent system that includes THF or methyl-THF (e.g.,2-MeTHF).

Synthetic Schemes

In this scheme, compounds of Formula 1 and 2 undergo palladium-catalyzedcross-coupling to generate the compound of Formula 3 in step x). Thecompound of Formula 3 undergoes recrystallization, in step xi), whereinit is converted to the tosylate salt 3-TsOH. The tosylate salt 3-TsOHundergoes deprotection using an inorganic base (e.g., LiOH) and work upin step ii) to generate the compound of Formula I (Free Form).

In another embodiment, the compound of Formula 2 and pharmaceuticallyacceptable salts thereof are prepared according to Scheme 2, below.

In step via), the compounds of Formula 8 and 9 undergo a nucleophilicsubstitution reaction to generate the compound of Formula 2, wherein X¹is defined above.

Some embodiments comprise the step of:

via) reacting a compound of Formula 8 with a compound of Formula 9,

in the presence of a base and an organic solvent to generate a mixturecomprising a compound of Formula 2 and a compound of Formula 10:

Some of these embodiments further comprising the steps of:

vii) reacting the mixture comprising the compound of Formula 2 and thecompound of Formula 10 with HCl in the presence of an organic solvent togenerate a mixture of hydrochloride salts of the compound of Formula 2and the compound of Formula 10; and

viii) recrystalizing the mixture of the hydrochloride salts of thecompound of Formula 2 and the compound of Formula 10 to generate thehydrochloride salt of the compound of Formula 2.

Some alternative embodiments comprise the steps of:

vib) reacting a compound of Formula 8 with a compound of Formula 9 inthe presence of a solvent and a base to generate the compound Formula 2

In some embodiments, the base of step vib) is an inorganic base selectedfrom tripotassium phosphate, dipotassium hydrogen phosphate, dipotassiumcarbonate, disodium carbonate, trisodium phosphate, disodium hydrogenphosphate, or any combination thereof.

In some embodiments, the solvent of step vib) comprises water. And, insome embodiments, the solvent of step vib) further comprises an alcoholselected from methanol, ethanol, propanol, iso-propanol, butanol,tert-butanol, or any combination thereof.

In some embodiments, the reaction of step vib) is performed at atemperature of from about 50° C. to about 100° C. For example, thereaction of step vib) is performed at a temperature of from about 60° C.to about 80° C.

The compound of Formula 9 and pharmaceutically acceptable salts arecommercially available.

In one specific embodiment, the step i) of the reaction of the compoundof Formula 1 with the compound of Formula 2 is performed in the presenceof a Pd-AmPhos catalyst and a base. In some embodiments, the step i) ofthe reaction of the compound of Formula 1 or a pharmaceuticallyacceptable salt thereof with the compound of Formula 2 is performed in asolvent system that includes water and an organic solvent selected from2-methyl THF or THF, or a combination thereof. In yet another specificembodiment, the deprotection step ii) comprises treating the compound ofFormula 3 or a pharmaceutically acceptable salt thereof with aninorganic hydroxide selected from the group consisting of LiOH, NaOH,and KOH. In yet another specific embodiment, the deprotection step ii)comprises treating the compound of Formula 3 or a pharmaceuticallyacceptable salt thereof with LiOH in a solvent system that includes THF.In yet another specific embodiment, the palladium catalyst of the step(f) of the reaction of the compound of Formula 4-2 or a pharmaceuticallyacceptable salt thereof with acetaldehyde includes a mixture ofbis(dibenzylideneacetone) palladium and a tertiary phosphine ligand,PR₃, wherein R is C₁₋₆ alkyl or C₅₋₆ cycloalkyl. In yet another specificembodiment, the tertiary phosphine ligand includes P(^(t)Bu)₃.

In yet another embodiment, the methods of the invention further employtreating the compound of Formula I, after the de-protecting step ii),with a solvent system comprising an organic solvent and water to form amonohydrate of the compound of Formula I. In one specific embodiment,the organic solvent of the solvent system comprises one or more organicsolvents independently selected from Class II organic solvents selectedfrom the group consisting of: chlorobenzene, cyclohexane,1,2-dichloroethene, dichloromethane, 1,2-dimethoxyethane,N,N-dimentylacetamide, N,N-dimethylformamide, 1,4-dioxane,2-ethoxyethanol, formamide, hexane, 2-methoxyethanol, methylbutylketone, methylcyclohexane, N-methylpyrrolidone, nitromethane, pyridine,sulfolane, tetrahydrofuran (THF), tetralin, tolune,1,1,2-trichloroethene and xylene, or Class III organic solvents selectedfrom the group consisting of: acetic acid, acetone, anisole, 1-butanol,2-butanol, butyl acetate, tert-butylmethyl ether, cumene, heptane,isobutyl acetate, isopropyl acetate, methyl acetate, 3-methyl-1-butanol,methylethyl ketone, methylisobutyl ketone, 2-methyl-1-propanol, ethylacetate, ethyl ether, ethyl formate, pentane, 1-pentanol, 1-propanol,2-propanol and propyl acetate. In another specific embodiment, theorganic solvents of the solvent system are selected from the groupconsisting of: chlorobenzene, cyclohexane, 1,2-dichloroethane,dichloromethane, 1,2-dimethoxyethane, formamide, hexane,2-methoxyethanol, methylbutyl ketone, methylcyclohexane, nitromethane,tetralin, xylene, toluene, 1,1,2-trichloroethane, acetone, anisole,1-butanol, 2-butanol, butyl acetate, t-butylmethylether, cumene,ethanol, ethyl acetate, ethyl ether, ethyl formate, heptane, isobutylacetate, isopropyl acetate, methyl acetate, 3-methyl-1-butanol,methylethyl ketone, 2-methy-1-propanol, pentane, 1-propanol, 1-pentanol,2-propanol, propyl acetate, tetrahydrofuran, and methyl tetrahydrofuran.In another specific embodiment, the organic solvents of the solventsystem are selected from the group consisting of: 2-ethoxyethanol,ethyleneglycol, methanol, 2-methoxyethanol, 1-butanol, 2-butanol,3-methyl-1-butanol, 2-methyl-1-propanol, ethanol, 1-pentanol,1-propanol, 2-propanol, methylbutyl ketone, acetone, methylethyl ketone,methylisobutyl ketone, butyl acetate, isobutyl acetate, isopropylacetate, methyl acetate, ethyl acetate, propyl acetate, pyridine,toluene, and xylene. In yet another specific embodiment, the solventsystem includes water and acetone, or water and isopropanol.

In some embodiments, the monohydrate (Hydrate 2) of the compound ofFormula Ia is characterized by one or more peaks corresponding to2-theta values measured in degrees of 6.9±0.2, 7.9±0.2, 13.8±0.2,15.9±0.2, 20.9±0.2, and 23.4±0.2 in an X-ray powder diffraction pattern.In some embodiments, this monohydrate is further characterized by one ormore peaks corresponding to 2-theta values measured in degrees of17.1±0.2, 18.6±0.2, 22.1±0.2 and 29.2±0.2 in an X-ray powder diffractionpattern. In other embodiments, the monohydrate of this compound ischaracterized by a ¹³C SSNMR spectrum of 178.5 ppm, 137.2 ppm, 126.8ppm, 107.0 ppm, and 35.3 ppm. In some embodiments, Hydrate 2 of Compound(1) is further characterized by a ¹³C SSNMR spectrum of 27.1 ppm.

Specific exemplary conditions suitable for each step of Schemes 1-4D, asprovided in the Examples below, may be independently employed in themethods of the invention.

The compounds described herein are defined herein by their chemicalstructures and/or chemical names. Where a compound is referred to byboth a chemical structure and a chemical name, and the chemicalstructure and chemical name conflict, the chemical structure isdeterminative of the compound's identity.

It will be appreciated by those skilled in the art that in the processesof the present invention certain functional groups such as hydroxyl oramino groups in the starting reagents or intermediate compounds may needto be protected by protecting groups. Thus, the preparation of thecompounds described above may involve, at various stages, the additionand removal of one or more protecting groups. The protection anddeprotection of functional groups is described in “Protective Groups inOrganic Chemistry.” edited by J. W. F. McOmie, Plenum Press (1973) and“Protective Groups in Organic Synthesis”, 3rd edition, T. W. Greene andP. G. M. Wuts, Wiley Interscience, and “Protecting Groups”, 3rd edition,P. J. Kocienski, Thieme (2005).

Selection of substituents and combinations of substituents envisioned bythis invention are those that result in the formation of stable orchemically feasible compounds. The term “stable”, as used herein, refersto compounds that are not substantially altered when subjected toconditions to allow for their production, detection, and, specifically,their recovery, purification, and use for one or more of the purposesdisclosed herein. In some embodiments, a stable compound or chemicallyfeasible compound is one that is not substantially altered when kept ata temperature of 40° C. or less, in the absence of moisture or otherchemically reactive conditions, for at least a week. Only those choicesand combinations of substituents that result in a stable structure arecontemplated. Such choices and combinations will be apparent to those ofordinary skill in the art and may be determined without undueexperimentation.

The term “protecting group” and “protective group” as used herein, areinterchangeable and refer to an agent used to temporarily block one ormore desired functional groups in a compound with multiple reactivesites. In certain embodiments, a protecting group has one or more, orspecifically all, of the following characteristics: a) is addedselectively to a functional group in good yield to give a protectedsubstrate that is b) stable to reactions occurring at one or more of theother reactive sites; and c) is selectively removable in good yield byreagents that do not attack the regenerated, deprotected functionalgroup. As would be understood by one skilled in the art, in some cases,the reagents do not attack other reactive groups in the compound. Inother cases, the reagents may also react with other reactive groups inthe compound. Examples of protecting groups are detailed in Greene, T.W., Wuts, P. G in “Protective Groups in Organic Synthesis”, ThirdEdition, John Wiley & Sons, New York: 1999 (and other editions of thebook), the entire contents of which are hereby incorporated byreference. The term “nitrogen protecting group”, as used herein, refersto an agent used to temporarily block one or more desired nitrogenreactive sites in a multifunctional compound. Preferred nitrogenprotecting groups also possess the characteristics exemplified for aprotecting group above, and certain exemplary nitrogen protecting groupsare also detailed in Chapter 7 in Greene, T. W., Wuts, P. G in“Protective Groups in Organic Synthesis”, Third Edition, John Wiley &Sons, New York: 1999, the entire contents of which are herebyincorporated by reference.

As used herein, the term “displaceable moiety” or “leaving group” refersto a group that is associated with an aliphatic or aromatic group asdefined herein and is subject to being displaced by nucleophilic attackby a nucleophile.

Unless otherwise indicated, structures depicted herein are also meant toinclude all isomeric (e.g., enantiomeric, diastereomeric, cis-trans,conformational, and rotational) forms of the structure. For example, theR and S configurations for each asymmetric center, (Z) and (E) doublebond isomers, and (Z) and (E) conformational isomers are included inthis invention, unless only one of the isomers is drawn specifically. Aswould be understood to one skilled in the art, a substituent can freelyrotate around any rotatable bonds. For example, a substituent drawn as

also represents

Therefore, single stereochemical isomers as well as enantiomeric,diastereomeric, cis/trans, conformational, and rotational mixtures ofthe present compounds are within the scope of the invention.

Unless otherwise indicated, all tautomeric forms of the compounds of theinvention are within the scope of the invention.

Additionally, unless otherwise indicated, structures depicted herein arealso meant to include compounds that differ only in the presence of oneor more isotopically enriched atoms. For example, compounds having thepresent structures except for the replacement of hydrogen by deuteriumor tritium, or the replacement of a carbon by a ¹³C- or ¹⁴C-enrichedcarbon are within the scope of this invention. Such compounds areuseful, for example, as analytical tools or probes in biological assays.Such compounds, especially deuterium analogs, can also betherapeutically useful.

It will be appreciated by those skilled in the art that the compounds inaccordance with the present invention can contain a chiral center. Thecompounds of formula may thus exist in the form of two different opticalisomers (i.e. (+) or (−) enantiomers). All such enantiomers and mixturesthereof including racemic mixtures are included within the scope of theinvention. The single optical isomer or enantiomer can be obtained bymethod well known in the art, such as chiral HPLC, enzymatic resolutionand chiral auxiliary.

In one embodiment, the compounds in accordance with the presentinvention are provided in the form of a single enantiomer at least 95%,at least 97% and at least 99% free of the corresponding enantiomer.

In a further embodiment, the compounds in accordance with the presentinvention are in the form of the (+) enantiomer at least 95% free of thecorresponding (−) enantiomer.

In a further embodiment, the compounds in accordance with the presentinvention are in the form of the (+) enantiomer at least 97% free of thecorresponding (−) enantiomer.

In a further embodiment, the compounds in accordance with the presentinvention are in the form of the (+) enantiomer at least 99% free of thecorresponding (−) enantiomer.

In a further embodiment, the compounds in accordance with the presentinvention are in the form of the (−) enantiomer at least 95% free of thecorresponding (+) enantiomer.

In a further embodiment, the compounds in accordance with the presentinvention are in the form of the (−) enantiomer at least 97% free of thecorresponding (+) enantiomer.

In a further embodiment the compounds in accordance with the presentinvention are in the form of the (−) enantiomer at least 99% free of thecorresponding (+) enantiomer.

IV. USES OF COMPOUNDS

The compounds disclosed herein can be used for inhibiting thereplication of influenza viruses in a biological sample or in a patient,for reducing the amount of influenza viruses (reducing viral titer) in abiological sample or in a patient, and for treating influenza in apatient. In one embodiment, the present invention is generally relatedto the use of the compounds disclosed herein (e.g., in pharmaceuticallyacceptable compositions) for any of the uses specified above.

In yet another embodiment, the compounds disclosed herein can be used toreduce viral titre in a biological sample (e.g. an infected cellculture) or in humans (e.g. lung viral titre in a patient).

The terms “influenza virus mediated condition”, “influenza infection”,or “Influenza”, as used herein, are used interchangeable to mean thedisease caused by an infection with an influenza virus.

Influenza is an infectious disease that affects birds and mammals causedby influenza viruses. Influenza viruses are RNA viruses of the familyOrthomyxoviridae, which comprises five genera: Influenza virus A,Influenza virus B, Influenza virus C, ISA virus and Thogoto virus.Influenza virus A genus has one species, influenza A virus which can besubdivided into different serotypes based on the antibody response tothese viruses: H1N1, H2N2, H3N2, H5N1, H7N7, H1N2, H9N2, H7N2, H7N3 andH10N7. Additional examples of influenza A virus include H3N8 and H7N9.Influenza virus B genus has one species, influenza B virus. Influenza Balmost exclusively infects humans and is less common than influenza A.Influenza virus C genus has one species, Influenza virus C virus, whichinfects humans and pigs and can cause severe illness and localepidemics. However, Influenza virus C is less common than the othertypes and usually seems to cause mild disease in children.

In some embodiments of the invention, influenza or influenza viruses areassociated with Influenza virus A or B. In some embodiments of theinvention, influenza or influenza viruses are associated with Influenzavirus A. In some specific embodiments of the invention, Influenza virusA is H1N1, H2N2, H3N2 or H5N1. In some specific embodiments of theinvention, Influenza virus A is H1N1, H3N2, H3N8, H5N1, and H7N9. Insome specific embodiments of the invention, Influenza virus A is H1N1,H3N2, H3N8, and H5N1.

In humans, common symptoms of influenza are chills, fever, pharyngitis,muscle pains, severe headache, coughing, weakness, and generaldiscomfort. In more serious cases, influenza causes pneumonia, which canbe fatal, particularly in young children and the elderly. Although it isoften confused with the common cold, influenza is a much more severedisease and is caused by a different type of virus. Influenza canproduce nausea and vomiting, especially in children, but these symptomsare more characteristic of the unrelated gastroenteritis, which issometimes called “stomach flu” or “24-hour flu”.

Symptoms of influenza can start quite suddenly one to two days afterinfection. Usually the first symptoms are chills or a chilly sensation,but fever is also common early in the infection, with body temperaturesranging from 38° C.-39° C. (approximately 100° F.-103° F.). Many peopleare so ill that they are confined to bed for several days, with achesand pains throughout their bodies, which are worse in their backs andlegs. Symptoms of influenza may include: body aches, especially jointsand throat, extreme coldness and fever, fatigue, headache, irritatedwatering eyes, reddened eyes, skin (especially face), mouth, throat andnose, abdominal pain (in children with influenza B). Symptoms ofinfluenza are non-specific, overlapping with many pathogens(“influenza-like illness). Usually, laboratory data is needed in orderto confirm the diagnosis.

The terms, “disease”, “disorder”, and “condition” may be usedinterchangeably here to refer to an influenza virus mediated medical orpathological condition.

As used herein, the terms “subject” and “patient” are usedinterchangeably. The terms “subject” and “patient” refer to an animal(e.g., a bird such as a chicken, quail or turkey, or a mammal),specifically a “mammal” including a non-primate (e.g., a cow, pig,horse, sheep, rabbit, guinea pig, rat, cat, dog, and mouse) and aprimate (e.g., a monkey, chimpanzee and a human), and more specificallya human. In one embodiment, the subject is a non-human animal such as afarm animal (e.g., a horse, cow, pig or sheep), or a pet (e.g., a dog,cat, guinea pig or rabbit). In a preferred embodiment, the subject is a“human”.

The term “biological sample”, as used herein, includes, withoutlimitation, cell cultures or extracts thereof; biopsied materialobtained from a mammal or extracts thereof; blood, saliva, urine, feces,semen, tears, or other body fluids or extracts thereof.

As used herein, “multiplicity of infection” or “MOI” is the ratio ofinfectious agents (e.g. phage or virus) to infection targets (e.g.cell). For example, when referring to a group of cells inoculated withinfectious virus particles, the multiplicity of infection or MOI is theratio defined by the number of infectious virus particles deposited in awell divided by the number of target cells present in that well.

As used herein the term “inhibition of the replication of influenzaviruses” includes both the reduction in the amount of virus replication(e.g. the reduction by at least 10%) and the complete arrest of virusreplication (i.e., 100% reduction in the amount of virus replication).In some embodiments, the replication of influenza viruses are inhibitedby at least 50%, at least 65%, at least 75%, at least 85%, at least 90%,or at least 95%.

Influenza virus replication can be measured by any suitable method knownin the art. For example, influenza viral titre in a biological sample(e.g. an infected cell culture) or in humans (e.g. lung viral titre in apatient) can be measured. More specifically, for cell based assays, ineach case cells are cultured in vitro, virus is added to the culture inthe presence or absence of a test agent, and after a suitable length oftime a virus-dependent endpoint is evaluated. For typical assays, theMadin-Darby canine kidney cells (MDCK) and the standard tissue cultureadapted influenza strain, A/Puerto Rico/8/34 can be used. A first typeof cell assay that can be used in the invention depends on death of theinfected target cells, a process called cytopathic effect (CPE), wherevirus infection causes exhaustion of the cell resources and eventuallysis of the cell. In the first type of cell assay, a low fraction ofcells in the wells of a microtiter plate are infected (typically 1/10 to1/1000), the virus is allowed to go through several rounds ofreplication over 48-72 hours, then the amount of cell death is measuredusing a decrease in cellular ATP content compared to uninfectedcontrols. A second type of cell assay that can be employed in theinvention depends on the multiplication of virus-specific RNA moleculesin the infected cells, with RNA levels being directly measured using thebranched-chain DNA hybridization method (bDNA). In the second type ofcell assay, a low number of cells are initially infected in wells of amicrotiter plate, the virus is allowed to replicate in the infectedcells and spread to additional rounds of cells, then the cells are lysedand viral RNA content is measured. This assay is stopped early, usuallyafter 18-36 hours, while all the target cells are still viable. ViralRNA is quantitated by hybridization to specific oligonucleotide probesfixed to wells of an assay plate, then amplification of the signal byhybridization with additional probes linked to a reporter enzyme.

As used herein a “viral titer (or titre)” is a measure of virusconcentration. Titer testing can employ serial dilution to obtainapproximate quantitative information from an analytical procedure thatinherently only evaluates as positive or negative. The titer correspondsto the highest dilution factor that still yields a positive reading; forexample, positive readings in the first 8 serial twofold dilutionstranslate into a titer of 1:256. A specific example is viral titer. Todetermine the titer, several dilutions will be prepared, such as 10⁻¹,10⁻², 10⁻³, 10⁻⁴, 10⁻⁵, 10⁻⁶, 10⁻⁷, 10⁻⁸. The lowest concentration ofvirus that still infects cells is the viral titer.

As used herein, the terms “treat”, “treatment”, and “treating” refer toboth therapeutic and prophylactic treatments. For example, therapeutictreatments includes the reduction or amelioration of the progression,severity and/or duration of influenza viruses mediated conditions, orthe amelioration of one or more symptoms (specifically, one or morediscernible symptoms) of influenza viruses mediated conditions,resulting from the administration of one or more therapies (e.g., one ormore therapeutic agents such as a compound or composition of theinvention). In specific embodiments, the therapeutic treatment includesthe amelioration of at least one measurable physical parameter of aninfluenza virus mediated condition. In other embodiments the therapeutictreatment includes the inhibition of the progression of an influenzavirus mediated condition, either physically by, e.g., stabilization of adiscernible symptom, physiologically by, e.g., stabilization of aphysical parameter, or both. In other embodiments the therapeutictreatment includes the reduction or stabilization of influenza virusesmediated infections. Antiviral drugs can be used in the communitysetting to treat people who already have influenza to reduce theseverity of symptoms and reduce the number of days that they are sick.

The term “chemotherapy” refers to the use of medications, e.g. smallmolecule drugs (rather than “vaccines”) for treating a disorder ordisease.

The terms “prophylaxis” or “prophylactic use” and “prophylactictreatment” as used herein, refer to any medical or public healthprocedure whose purpose is to prevent, rather than treat or cure adisease. As used herein, the terms “prevent”, “prevention” and“preventing” refer to the reduction in the risk of acquiring ordeveloping a given condition, or the reduction or inhibition of therecurrence or said condition in a subject who is not ill, but who hasbeen or may be near a person with the disease. The term“chemoprophylaxis” refers to the use of medications, e.g. small moleculedrugs (rather than “vaccines”) for the prevention of a disorder ordisease.

As used herein, prophylactic use includes the use in situations in whichan outbreak has been detected, to prevent contagion or spread of theinfection in places where a lot of people that are at high risk ofserious influenza complications live in close contact with each other(e.g. in a hospital ward, daycare center, prison, nursing home, etc.).It also includes the use among populations who require protection fromthe influenza but who either do not get protection after vaccination(e.g. due to weak immune system), or when the vaccine is unavailable tothem, or when they cannot get the vaccine because of side effects. Italso includes use during the two weeks following vaccination, sinceduring that time the vaccine is still ineffective. Prophylactic use mayalso include treating a person who is not ill with the influenza or notconsidered at high risk for complications, in order to reduce thechances of getting infected with the influenza and passing it on to ahigh-risk person in close contact with him (for instance, healthcareworkers, nursing home workers, etc.).

According to the US CDC, an influenza “outbreak” is defined as a suddenincrease of acute febrile respiratory illness (AFRI) occurring within a48 to 72 hour period, in a group of people who are in close proximity toeach other (e.g. in the same area of an assisted living facility, in thesame household, etc.) over the normal background rate or when anysubject in the population being analyzed tests positive for influenza.One case of confirmed influenza by any testing method is considered anoutbreak.

A “cluster” is defined as a group of three or more cases of AFRIoccurring within a 48 to 72 hour period, in a group of people who are inclose proximity to each other (e.g. in the same area of an assistedliving facility, in the same household, etc.).

As used herein, the “index case”, “primary case”, or “patient zero” isthe initial patient in the population sample of an epidemiologicalinvestigation. When used in general to refer to such patients inepidemiological investigations, the term is not capitalized. When theterm is used to refer to a specific person in place of that person'sname within a report on a specific investigation, the term iscapitalized as Patient Zero. Often scientists search for the index caseto determine how the disease spread and what reservoir holds the diseasein between outbreaks. Note that the index case is the first patient thatindicates the existence of an outbreak. Earlier cases may be found andare labeled primary, secondary, tertiary, etc.

In one embodiment, the methods of the invention are a preventative or“pre-emptive” measure to a patient, specifically a human, having apredisposition to complications resulting from infection by an influenzavirus. The term “pre-emptive” as used herein as for example inpre-emptive use, “pre-emptively”, etc., is the prophylactic use insituations in which an “index case” or an “outbreak” has been confirmed,in order to prevent the spread of infection in the rest of the communityor population group.

In another embodiment, the methods of the invention are applied as a“pre-emptive” measure to members of a community or population group,specifically humans, in order to prevent the spread of infection.

As used herein, an “effective amount” refers to an amount sufficient toelicit the desired biological response. In the present invention thedesired biological response is to inhibit the replication of influenzavirus, to reduce the amount of influenza viruses or to reduce orameliorate the severity, duration, progression, or onset of a influenzavirus infection, prevent the advancement of an influenza virusesinfection, prevent the recurrence, development, onset or progression ofa symptom associated with an influenza virus infection, or enhance orimprove the prophylactic or therapeutic effect(s) of another therapyused against influenza infections. The precise amount of compoundadministered to a subject will depend on the mode of administration, thetype and severity of the infection and on the characteristics of thesubject, such as general health, age, sex, body weight and tolerance todrugs. The skilled artisan will be able to determine appropriate dosagesdepending on these and other factors. When co-administered with otherantiviral agents, e.g., when co-administered with an anti-influenzamedication, an “effective amount” of the second agent will depend on thetype of drug used. Suitable dosages are known for approved agents andcan be adjusted by the skilled artisan according to the condition of thesubject, the type of condition(s) being treated and the amount of acompound described herein being used. In cases where no amount isexpressly noted, an effective amount should be assumed. For example, thecompounds disclosed herein can be administered to a subject in a dosagerange from between approximately 0.01 to 100 mg/kg body weight/day fortherapeutic or prophylactic treatment.

Generally, dosage regimens can be selected in accordance with a varietyof factors including the disorder being treated and the severity of thedisorder; the activity of the specific compound employed; the specificcomposition employed; the age, body weight, general health, sex and dietof the patient; the time of administration, route of administration, andrate of excretion of the specific compound employed; the renal andhepatic function of the subject; and the particular compound or saltthereof employed, the duration of the treatment; drugs used incombination or coincidental with the specific compound employed, andlike factors well known in the medical arts. The skilled artisan canreadily determine and prescribe the effective amount of the compoundsdescribed herein required to treat, to prevent, inhibit (fully orpartially) or arrest the progress of the disease.

Dosages of the compounds described herein can range from between 0.01mg/kg to 100 mg/kg body weight/day, 0.01 mg/kg to 50 mg/kg bodyweight/day, 0.1 mg/kg to 50 mg/kg body weight/day, or 1 mg/kg to 25mg/kg body weight/day. It is understood that the total amount per daycan be administered in a single dose or can be administered in multipledosing, such as twice a day (e.g., every 12 hours), three times a day(e.g., every 8 hours), or four times a day (e.g., every 6 hours).

In some embodiments, dosages of the compounds described herein (e.g.,the compound of Formula I and its pharmaceutically acceptable saltsthereof, including the various solid forms) are in a range of 100 mg to1,600 mg, such as 400 mg to 1,600 mg or 400 mg to 1,200 mg. Each dosecan be taken once a day (QD), twice per day (e.g., approximately every12 hours (BID)), or three times per day (e.g., approximately every 8hours (TID)). It is noted that any combinations of QD, BID, and TID canbe employed, as desired, such as BID on day 1, followed by QDthereafter.

In one specific embodiment, dosages of the compounds described hereinare from 400 mg to 1,600 mg, from 400 mg to 1,200 mg, or from 600 mg to1,200 mg once a day. In another specific embodiment, dosages of thecompounds described herein are from 400 mg to 1,600 mg, from 400 mg to1,200 mg, or from 300 mg to 900 mg twice a day. In yet another specificembodiment, dosages of the compounds described herein are from 400 mg to1,000 mg once a day. In yet another specific embodiment, dosages of thecompounds described herein are from 600 mg to 1,000 mg once a day. Inyet another specific embodiment, dosages of the compounds describedherein are from 600 mg to 800 mg once a day. In yet another specificembodiment, dosages of the compounds described herein are from 400 mg to800 mg twice a day (e.g., from 400 mg to 800 mg every 12 hours). In yetanother specific embodiment, dosages of the compounds described hereinare from 400 mg to 600 mg twice a day.

In some embodiments, a loading dosage regimen is employed. In onespecific embodiment, a loading dose of from 400 mg to 1,600 mg isemployed on day 1 of treatment. In another specific embodiment, aloading dose of from 600 mg to 1,600 mg is employed on day 1 oftreatment. In another specific embodiment, a loading dose of from 800 mgto 1,600 mg is employed on day 1 of treatment. In yet another specificembodiment, a loading dose of from 900 mg to 1,600 mg is employed on day1 of treatment. In yet another specific embodiment, a loading dose offrom 900 mg to 1,200 mg is employed on day 1 of treatment. In yetanother specific embodiment, a loading dose of 900 mg is employed on day1 of treatment. In yet another specific embodiment, a loading dose of1,000 mg is employed on day 1 of treatment. In yet another specificembodiment, a loading dose of 1,200 mg is employed on day 1 oftreatment.

In one specific embodiment, the dosage regimen of the compoundsdescribed herein employs a loading dosage of 600 mg to 1,600 mg on day 1and with a regular dosage of 300 mg to 1,200 mg for the rest of thetreatment duration. Each regular dose can be taken once a day, twice aday, or three times a day, or any combination thereof. In a furtherspecific embodiment, a loading dosage of 900 mg to 1,600 mg, such as 900mg, 1,200 mg, or 1,600 mg, is employed. In another further specificembodiment, a loading dosage of 900 mg to 1,200 mg, such as 900 mg or1,200 mg, is employed. In yet another further specific embodiment, aregular dosage of 400 mg to 1,200 mg, such as 400 mg, 600 mg, or 800 mg,is employed for the rest of the treatment duration. In yet anotherfurther specific embodiment, a regular dosage of 400 mg to 1,000 mg forthe rest of the treatment duration. In yet another further specificembodiment, a regular dosage of 400 mg to 800 mg is employed for therest of the treatment duration. In yet another further specificembodiment, a regular dosage of 300 mg to 900 mg twice a day isemployed. In yet another further specific embodiment, a regular dosageof 600 mg to 1,200 mg once a day is employed. In yet another furtherspecific embodiment, a regular dosage of 600 mg twice a day on day 2,followed by 600 mg once a day for the rest of the treatment duration.

For therapeutic treatment, the compounds described herein can beadministered to a patient within, for example, 48 hours (or within 40hours, or less than 2 days, or less than 1.5 days, or within 24 hours)of onset of symptoms (e.g., nasal congestion, sore throat, cough, aches,fatigue, headaches, and chills/sweats). Alternatively, for therapeutictreatment, the compounds described herein can be administered to apatient within, for example, 96 hours of onset of symptoms. Thetherapeutic treatment can last for any suitable duration, for example,for 3 days, 4 days, 5 days, 7 days, 10 days, 14 days, etc. Forprophylactic treatment during a community outbreak, the compoundsdescribed herein can be administered to a patient within, for example, 2days of onset of symptoms in the index case, and can be continued forany suitable duration, for example, for 7 days, 10 days, 14 days, 20days, 28 days, 35 days, 42 days, etc., up to the entire flu season. Aflu season is an annually-recurring time period characterized by theprevalence of outbreaks of influenza. Influenza activity can sometimesbe predicted and even tracked geographically. While the beginning ofmajor flu activity in each season varies by location, in any specificlocation these minor epidemics usually take 3-4 weeks to peak andanother 3-4 weeks to significantly diminish. Typically, Centers forDisease Control (CDC) collects, compiles and analyzes information oninfluenza activity year round in the United States and produces a weeklyreport from October through mid-May.

In one embodiment, the therapeutic treatment lasts for 1 day to anentire flu season. In one specific embodiment, the therapeutic treatmentlasts for 3 days to 14 days. In another specific embodiment, thetherapeutic treatment lasts for 5 days to 14 days. In another specificembodiment, the therapeutic treatment lasts for 3 days to 10 days. Inyet another specific embodiment, the therapeutic treatment lasts for 4days to 10 days. In yet another specific embodiment, the therapeutictreatment lasts for 5 days to 10 days. In yet another specificembodiment, the therapeutic treatment lasts for 4 days to 7 days (e.g.,4 days, 5 days, 6 days, or 7 days). In yet another specific embodiment,the therapeutic treatment lasts for 5 days to 7 days (e.g., 5 days, 6days, or 7 days). In one specific embodiment, the prophylactic treatmentlasts up to the entire flu season.

In one specific embodiment, the compounds described herein areadministered to a patient for 3 days to 14 days (e.g., 5 days to 14days) with a loading dosage of 900 mg to 1,600 mg on day 1 and with aregular dosage of 300 mg to 1,200 mg for the rest of the treatmentduration. In another specific embodiment, the compounds described hereinare administered to a patient for 3 days to 14 days (e.g., 5 days to 14days) with a loading dosage of 900 mg to 1,200 mg on day 1 and with aregular dosage of 400 mg to 1,000 mg for the rest of the treatmentduration. In yet another specific embodiment, the compounds describedherein are administered to a patient for 3 days to 14 days (e.g., 5 daysto 14 days) with a loading dosage of 900 mg to 1,200 mg on day 1 andwith a regular dosage of 400 mg to 800 mg for the rest of the treatmentduration. In yet another specific embodiment, the compounds describedherein are administered to a patient for 3 days to 14 days (e.g., 5 daysto 14 days) with a loading dosage of 900 mg to 1,200 mg on day 1 andwith a regular dosage of 400 mg to 800 mg for the rest of the treatmentduration. Each dose can be taken once a day, twice a day, or three timesa day, or any combination thereof.

In one specific embodiment, the compounds described herein areadministered to a patient for 3 days to 14 days with a loading dosage of900 mg to 1,600 mg on day 1 and with a regular dosage of 600 mg to 1,000mg once a day for the rest of the treatment duration. In anotherspecific embodiment, the compounds described herein are administered toa patient for 3 days to 14 days with a loading dosage of 900 mg to 1,200mg on day 1 and with a regular dosage of 600 mg to 800 mg (e.g., 600 mg,650 mg, 700 mg, 750 mg, or 800 mg) once a day for the rest of thetreatment duration. In some embodiments, the treatment duration is for 4days to 10 days, 5 days to 10 days, or 5 days to 7 days.

In one specific embodiment, the compounds described herein areadministered to a patient for 3 days to 14 days with a loading dosage of900 mg to 1,600 mg on day 1 and with a regular dosage of 400 mg to 800mg twice a day for the rest of the treatment duration. In anotherspecific embodiment, the compounds described herein are administered toa patient for 3 days to 14 days with a loading dosage of 900 mg to 1,200mg on day 1 and with a regular dosage of 400 mg to 600 mg (e.g., 400 mg,450 mg, 500 mg, 550 mg, or 600 mg) twice a day for the rest of thetreatment duration. In some embodiments, the duration is for 4 days to10 days, 5 days to 10 days, or 5 days to 7 days.

In one specific embodiment, the compounds described herein areadministered to a patient for 4 days or 5 days with a loading dosage of900 mg to 1,200 mg (e.g., 900 mg or 1,200 mg) on day 1 and with aregular dosage of 400 mg to 600 mg (e.g., 400 mg or 600 mg) twice a dayfor the rest of the treatment duration (e.g., days 2 through 4, or days2 through 5). In another specific embodiment, the compounds describedherein are administered to a patient for 4 days or 5 days with a loadingdosage of 900 mg to 1,200 mg (e.g., 900 mg or 1,200 mg) on day 1 andwith a regular dosage of 600 mg to 800 mg (e.g., 600 mg or 800 mg) oncea day for the rest of the treatment duration.

Various types of administration methods can be employed in theinvention, and are described in detail below under the section entitled“Administration Methods”.

V. COMBINATION THERAPY

An effective amount can be achieved in the method or pharmaceuticalcomposition of the invention employing a compound of the invention(including a pharmaceutically acceptable salt or solvate (e.g., ahydrate)) alone or in combination with an additional suitabletherapeutic agent, for example, an antiviral agent or a vaccine. When a“combination therapy” is employed, an effective amount can be achievedusing a first amount of a compound of the invention and a second amountof an additional suitable therapeutic agent (e.g. an antiviral agent orvaccine).

In another embodiment of this invention, a compound of the invention andthe additional therapeutic agent, are each administered in an effectiveamount (i.e., each in an amount which would be therapeutically effectiveif administered alone). In another embodiment, a compound of theinvention and the additional therapeutic agent, are each administered inan amount which alone does not provide a therapeutic effect (asub-therapeutic dose). In yet another embodiment, a compound of theinvention can be administered in an effective amount, while theadditional therapeutic agent is administered in a sub-therapeutic dose.In still another embodiment, a compound of the invention can beadministered in a sub-therapeutic dose, while the additional therapeuticagent, for example, a suitable cancer-therapeutic agent is administeredin an effective amount.

As used herein, the terms “in combination” or “co-administration” can beused interchangeably to refer to the use of more than one therapy (e.g.,one or more prophylactic and/or therapeutic agents). The use of theterms does not restrict the order in which therapies (e.g., prophylacticand/or therapeutic agents) are administered to a subject.

Co-administration encompasses administration of the first and secondamounts of the compounds of the co-administration in an essentiallysimultaneous manner, such as in a single pharmaceutical composition, forexample, capsule or tablet having a fixed ratio of first and secondamounts, or in multiple, separate capsules or tablets for each. Inaddition, such co-administration also encompasses use of each compoundin a sequential manner in either order.

In one embodiment, the present invention is directed to methods ofcombination therapy for inhibiting Flu viruses replication in biologicalsamples or patients, or for treating or preventing Influenza virusinfections in patients using the compounds described herein.Accordingly, pharmaceutical compositions of the invention also includethose comprising an inhibitor of Flu virus replication of this inventionin combination with an anti-viral compound exhibiting anti-Influenzavirus activity.

Methods of use of the compounds described herein and compositions of theinvention also include combination of chemotherapy with a compound orcomposition of the invention, or with a combination of a compound orcomposition of this invention with another anti-viral agent andvaccination with a Flu vaccine.

When co-administration involves the separate administration of the firstamount of a compound of the invention and a second amount of anadditional therapeutic agent, the compounds are administeredsufficiently close in time to have the desired therapeutic effect. Forexample, the period of time between each administration which can resultin the desired therapeutic effect, can range from minutes to hours andcan be determined taking into account the properties of each compoundsuch as potency, solubility, bioavailability, plasma half-life andkinetic profile. For example, a compound of the invention and the secondtherapeutic agent can be administered in any order within 24 hours ofeach other, within 16 hours of each other, within 8 hours of each other,within 4 hours of each other, within 1 hour of each other or within 30minutes of each other.

More, specifically, a first therapy (e.g., a prophylactic or therapeuticagent such as a compound of the invention) can be administered prior to(e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeksbefore), concomitantly with, or subsequent to (e.g., 5 minutes, 15minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks,4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks after) theadministration of a second therapy (e.g., a prophylactic or therapeuticagent such as an anti-cancer agent) to a subject.

It is understood that the method of co-administration of a first amountof a compound of the invention and a second amount of an additionaltherapeutic agent can result in an enhanced or synergistic therapeuticeffect, wherein the combined effect is greater than the additive effectthat would result from separate administration of the first amount of acompound of the invention and the second amount of an additionaltherapeutic agent.

As used herein, the term “synergistic” refers to a combination of acompound of the invention and another therapy (e.g., a prophylactic ortherapeutic agent), which is more effective than the additive effects ofthe therapies. A synergistic effect of a combination of therapies (e.g.,a combination of prophylactic or therapeutic agents) can permit the useof lower dosages of one or more of the therapies and/or less frequentadministration of said therapies to a subject. The ability to utilizelower dosages of a therapy (e.g., a prophylactic or therapeutic agent)and/or to administer said therapy less frequently can reduce thetoxicity associated with the administration of said therapy to a subjectwithout reducing the efficacy of said therapy in the prevention,management or treatment of a disorder. In addition, a synergistic effectcan result in improved efficacy of agents in the prevention, managementor treatment of a disorder. Finally, a synergistic effect of acombination of therapies (e.g., a combination of prophylactic ortherapeutic agents) may avoid or reduce adverse or unwanted side effectsassociated with the use of either therapy alone.

When the combination therapy using the compounds of the presentinvention is in combination with a Flu vaccine, both therapeutic agentscan be administered so that the period of time between eachadministration can be longer (e.g. days, weeks or months).

The presence of a synergistic effect can be determined using suitablemethods for assessing drug interaction. Suitable methods include, forexample, the Sigmoid-Emax equation (Holford, N. H. G. and Scheiner, L.B., Clin. Pharmacokinet. 6: 429-453 (1981)), the equation of Loeweadditivity (Loewe, S. and Muischnek, H., Arch. Exp. Pathol Pharmacol.114: 313-326 (1926)) and the median-effect equation (Chou, T. C. andTalalay, P., Adv. Enzyme Regul. 22: 27-55 (1984)). Each equationreferred to above can be applied with experimental data to generate acorresponding graph to aid in assessing the effects of the drugcombination. The corresponding graphs associated with the equationsreferred to above are the concentration-effect curve, isobologram curveand combination index curve, respectively.

Specific examples that can be co-administered with a compound describedherein include neuraminidase inhibitors, such as oseltamivir (Tamiflu®)and Zanamivir (Rlenza®), viral ion channel (M2 protein) blockers, suchas amantadine (Symmetrel®) and rimantadine (Flumadine®), and antiviraldrugs described in WO 2003/015798, including T-705 under development byToyama Chemical of Japan. (See also Ruruta et al., Antiviral Research,82: 95-102 (2009), “T-705 (flavipiravir) and related compounds: Novelbroad-spectrum inhibitors of RNA viral infections”). In someembodiments, the compounds described herein can be co-administered witha traditional influenza vaccine. In some embodiments, the compoundsdescribed herein can be co-administered with Zanamivir. In someembodiments, the compounds described herein can be co-administered withoseltamivir. In some embodiments, the compounds described herein can beco-administered with T-705. In some embodiments, the compounds describedherein can be co-administered with amantadine or rimantadine.Oseltamivir can be administered in a dosage regimen specified in itslabel. In some specific embodiments, it is administered 75 mg twice aday, or 150 mg once a day.

VI. PHARMACEUTICAL COMPOSITIONS

The compounds described herein can be formulated into pharmaceuticalcompositions that further comprise a pharmaceutically acceptablecarrier, diluent, adjuvant or vehicle. In one embodiment, the presentinvention relates to a pharmaceutical composition comprising a compoundof the invention described above, and a pharmaceutically acceptablecarrier, diluent, adjuvant or vehicle. In one embodiment, the presentinvention is a pharmaceutical composition comprising an effective amountof a compound of the present invention or a pharmaceutically acceptablesalt thereof and a pharmaceutically acceptable carrier, diluent,adjuvant or vehicle. Pharmaceutically acceptable carriers include, forexample, pharmaceutical diluents, excipients or carriers suitablyselected with respect to the intended form of administration, andconsistent with conventional pharmaceutical practices.

An “effective amount” includes a “therapeutically effective amount” anda “prophylactically effective amount”. The term “therapeuticallyeffective amount” refers to an amount effective in treating and/orameliorating an influenza virus infection in a patient infected withinfluenza. The term “prophylactically effective amount” refers to anamount effective in preventing and/or substantially lessening thechances or the size of influenza virus infection outbreak. Specificexamples of effective amounts are described above in the sectionentitled Uses of Disclosed Compounds.

A pharmaceutically acceptable carrier may contain inert ingredientswhich do not unduly inhibit the biological activity of the compounds.The pharmaceutically acceptable carriers should be biocompatible, e.g.,non-toxic, non-inflammatory, non-immunogenic or devoid of otherundesired reactions or side-effects upon the administration to asubject. Standard pharmaceutical formulation techniques can be employed.

The pharmaceutically acceptable carrier, adjuvant, or vehicle, as usedherein, includes any and all solvents, diluents, or other liquidvehicle, dispersion or suspension aids, surface active agents, isotonicagents, thickening or emulsifying agents, preservatives, solid binders,lubricants and the like, as suited to the particular dosage formdesired. Remington's Pharmaceutical Sciences, Sixteenth Edition, E. W.Martin (Mack Publishing Co., Easton, Pa., 1980) discloses variouscarriers used in formulating pharmaceutically acceptable compositionsand known techniques for the preparation thereof. Except insofar as anyconventional carrier medium is incompatible with the compounds describedherein, such as by producing any undesirable biological effect orotherwise interacting in a deleterious manner with any othercomponent(s) of the pharmaceutically acceptable composition, its use iscontemplated to be within the scope of this invention. As used herein,the phrase “side effects” encompasses unwanted and adverse effects of atherapy (e.g., a prophylactic or therapeutic agent). Side effects arealways unwanted, but unwanted effects are not necessarily adverse. Anadverse effect from a therapy (e.g., prophylactic or therapeutic agent)might be harmful or uncomfortable or risky. Side effects include, butare not limited to fever, chills, lethargy, gastrointestinal toxicities(including gastric and intestinal ulcerations and erosions), nausea,vomiting, neurotoxicities, nephrotoxicities, renal toxicities (includingsuch conditions as papillary necrosis and chronic interstitialnephritis), hepatic toxicities (including elevated serum liver enzymelevels), myelotoxicities (including leukopenia, myelosuppression,thrombocytopenia and anemia), dry mouth, metallic taste, prolongation ofgestation, weakness, somnolence, pain (including muscle pain, bone painand headache), hair loss, asthenia, dizziness, extra-pyramidal symptoms,akathisia, cardiovascular disturbances and sexual dysfunction.

Some examples of materials which can serve as pharmaceuticallyacceptable carriers include, but are not limited to, ion exchangers,alumina, aluminum stearate, lecithin, serum proteins (such as humanserum albumin), buffer substances (such as twin 80, phosphates, glycine,sorbic acid, or potassium sorbate), partial glyceride mixtures ofsaturated vegetable fatty acids, water, salts or electrolytes (such asprotamine sulfate, disodium hydrogen phosphate, potassium hydrogenphosphate, sodium chloride, or zinc salts), colloidal silica, magnesiumtrisilicate, polyvinyl pyrrolidone, polyacrylates, waxes,polyethylene-polyoxypropylene-block polymers, methylcellulose,hydroxypropyl methylcellulose, wool fat, sugars such as lactose, glucoseand sucrose; starches such as corn starch and potato starch; celluloseand its derivatives such as sodium carboxymethyl cellulose, ethylcellulose and cellulose acetate; powdered tragacanth; malt; gelatin;talc; excipients such as cocoa butter and suppository waxes; oils suchas peanut oil, cottonseed oil; safflower oil; sesame oil; olive oil;corn oil and soybean oil; glycols; such a propylene glycol orpolyethylene glycol; esters such as ethyl oleate and ethyl laurate;agar; buffering agents such as magnesium hydroxide and aluminumhydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer'ssolution; ethyl alcohol, and phosphate buffer solutions, as well asother non-toxic compatible lubricants such as sodium lauryl sulfate andmagnesium stearate, as well as coloring agents, releasing agents,coating agents, sweetening, flavoring and perfuming agents,preservatives and antioxidants can also be present in the composition,according to the judgment of the formulator.

VII. ADMINISTRATION METHODS

The compounds and pharmaceutically acceptable compositions describedabove can be administered to humans and other animals orally, rectally,parenterally, intracisternally, intravaginally, intraperitoneally,topically (as by powders, ointments, or drops), bucally, as an oral ornasal spray, or the like, depending on the severity of the infectionbeing treated.

Liquid dosage forms for oral administration include, but are not limitedto, pharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active compounds,the liquid dosage forms may contain inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor, and sesame oils),glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fattyacid esters of sorbitan, and mixtures thereof. Besides inert diluents,the oral compositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

The injectable formulations can be sterilized, for example, byfiltration through a bacterial-retaining filter, or by incorporatingsterilizing agents in the form of sterile solid compositions which canbe dissolved or dispersed in sterile water or other sterile injectablemedium prior to use.

In order to prolong the effect of a compound described herein, it isoften desirable to slow the absorption of the compound from subcutaneousor intramuscular injection. This may be accomplished by the use of aliquid suspension of crystalline or amorphous material with poor watersolubility. The rate of absorption of the compound then depends upon itsrate of dissolution that, in turn, may depend upon crystal size andcrystalline form. Alternatively, delayed absorption of a parenterallyadministered compound form is accomplished by dissolving or suspendingthe compound in an oil vehicle. Injectable depot forms are made byforming microencapsule matrices of the compound in biodegradablepolymers such as polylactide-polyglycolide. Depending upon the ratio ofcompound to polymer and the nature of the particular polymer employed,the rate of compound release can be controlled. Examples of otherbiodegradable polymers include poly(orthoesters) and poly(anhydrides).Depot injectable formulations are also prepared by entrapping thecompound in liposomes or microemulsions that are compatible with bodytissues.

Compositions for rectal or vaginal administration are specificallysuppositories which can be prepared by mixing the compounds describedherein with suitable non-irritating excipients or carriers such as cocoabutter, polyethylene glycol or a suppository wax which are solid atambient temperature but liquid at body temperature and therefore melt inthe rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like. The solid dosage forms of tablets, dragees, capsules, pills,and granules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes. Solid compositions of a similartype may also be employed as fillers in soft and hard-filled gelatincapsules using such excipients as lactose or milk sugar as well as highmolecular weight polethylene glycols and the like.

The active compounds can also be in microencapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositions thatcan be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compounddescribed herein include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, eardrops, and eye drops are also contemplated asbeing within the scope of this invention. Additionally, the presentinvention contemplates the use of transdermal patches, which have theadded advantage of providing controlled delivery of a compound to thebody. Such dosage forms can be made by dissolving or dispensing thecompound in the proper medium. Absorption enhancers can also be used toincrease the flux of the compound across the skin. The rate can becontrolled by either providing a rate controlling membrane or bydispersing the compound in a polymer matrix or gel.

The compositions described herein may be administered orally,parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir. The term “parenteral”as used herein includes, but is not limited to, subcutaneous,intravenous, intramuscular, intra-articular, intra-synovial,intrasternal, intrathecal, intrahepatic, intralesional and intracranialinjection or infusion techniques. Specifically, the compositions areadministered orally, intraperitoneally or intravenously.

Sterile injectable forms of the compositions described herein may beaqueous or oleaginous suspension. These suspensions may be formulatedaccording to techniques known in the art using suitable dispersing orwetting agents and suspending agents. The sterile injectable preparationmay also be a sterile injectable solution or suspension in a non-toxicparenterally-acceptable diluent or solvent, for example as a solution in1,3-butanediol. Among the acceptable vehicles and solvents that may beemployed are water, Ringer's solution and isotonic sodium chloridesolution. In addition, sterile, fixed oils are conventionally employedas a solvent or suspending medium. For this purpose, any bland fixed oilmay be employed including synthetic mono- or di-glycerides. Fatty acids,such as oleic acid and its glyceride derivatives are useful in thepreparation of injectables, as are natural pharmaceutically-acceptableoils, such as olive oil or castor oil, especially in theirpolyoxyethylated versions. These oil solutions or suspensions may alsocontain a long-chain alcohol diluent or dispersant, such ascarboxymethyl cellulose or similar dispersing agents which are commonlyused in the formulation of pharmaceutically acceptable dosage formsincluding emulsions and suspensions. Other commonly used surfactants,such as Tweens, Spans and other emulsifying agents or bioavailabilityenhancers which are commonly used in the manufacture of pharmaceuticallyacceptable solid, liquid, or other dosage forms may also be used for thepurposes of formulation.

The pharmaceutical compositions described herein may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, aqueous suspensions or solutions. In thecase of tablets for oral use, carriers commonly used include, but arenot limited to, lactose and corn starch. Lubricating agents, such asmagnesium stearate, are also typically added. For oral administration ina capsule form, useful diluents include lactose and dried cornstarch.When aqueous suspensions are required for oral use, the activeingredient is combined with emulsifying and suspending agents. Ifdesired, certain sweetening, flavoring or coloring agents may also beadded.

Alternatively, the pharmaceutical compositions described herein may beadministered in the form of suppositories for rectal administration.These can be prepared by mixing the agent with a suitable non-irritatingexcipient which is solid at room temperature but liquid at rectaltemperature and therefore will melt in the rectum to release the drug.Such materials include, but are not limited to, cocoa butter, beeswaxand polyethylene glycols.

The pharmaceutical compositions described herein may also beadministered topically, especially when the target of treatment includesareas or organs readily accessible by topical application, includingdiseases of the eye, the skin, or the lower intestinal tract. Suitabletopical formulations are readily prepared for each of these areas ororgans.

Topical application for the lower intestinal tract can be effected in arectal suppository formulation (see above) or in a suitable enemaformulation. Topically-transdermal patches may also be used.

For topical applications, the pharmaceutical compositions may beformulated in a suitable ointment containing the active componentsuspended or dissolved in one or more carriers. Carriers for topicaladministration of the compounds of this invention include, but are notlimited to, mineral oil, liquid petrolatum, white petrolatum, propyleneglycol, polyoxyethylene, polyoxypropylene compound, emulsifying wax andwater. Alternatively, the pharmaceutical compositions can be formulatedin a suitable lotion or cream containing the active components suspendedor dissolved in one or more pharmaceutically acceptable carriers.Suitable carriers include, but are not limited to, mineral oil, sorbitanmonostearate, polysorbate 60, cetyl esters wax, cetearyl alcohol, 2octyldodecanol, benzyl alcohol and water.

For ophthalmic use, the pharmaceutical compositions may be formulated asmicronized suspensions in isotonic, pH adjusted sterile saline, or,specifically, as solutions in isotonic, pH adjusted sterile saline,either with or without a preservative such as benzylalkonium chloride.Alternatively, for ophthalmic uses, the pharmaceutical compositions maybe formulated in an ointment such as petrolatum.

The pharmaceutical compositions may also be administered by nasalaerosol or inhalation. Such compositions are prepared according totechniques well-known in the art of pharmaceutical formulation and maybe prepared as solutions in saline, employing benzyl alcohol or othersuitable preservatives, absorption promoters to enhance bioavailability,fluorocarbons, and/or other conventional solubilizing or dispersingagents.

The compounds can be formulated in unit dosage form. The term “unitdosage form” refers to physically discrete units suitable as unitarydosage for subjects undergoing treatment, with each unit containing apredetermined quantity of active material calculated to produce thedesired therapeutic effect, optionally in association with a suitablepharmaceutical carrier. The unit dosage form can be for a single dailydose or one of multiple daily doses (e.g., 1 to 4 or more times perday). When multiple daily doses are used, the unit dosage form can bethe same or different for each dose.

VIII. EXAMPLES Example 1: Preparation of2-Amino-3-bromo-5-fluoropyridine (4-2a)

Procedure A:

To a slurry of 2-amino-5-fluoropyridine (6 kg, 53.6 mol) in water (24 L)at 14° C. was added over 10 minutes 48% hydrobromic acid (18.5 kg, 110mol). The reaction was exothermic and the temperature went up to 24° C.The mixture was re-cooled to 12° C. then bromine (9 kg, 56.3 mol) wasadded in nine portions over 50 minutes (exothermic, kept at 20° C.). Themixture was stirred at 22° C. overnight, and monitored by ¹HNMR of aquenched aliquot (quenched 5 drops in to mix of 1 ml 20% K₂CO₃, 0.3 ml10% Na₂S₂O₃ and 0.7 ml DCM. Organic layer evaporated and assayed). Themixture was cooled to 10° C. then quenched by addition of sodiumbisulfite (560 g, 5.4 mol) in water (2 L), and further cooled to 0° C.This mixture was added to a cold (−4° C.) mixture of DCM (18 L) and 5.4Msodium hydroxide (35 L, 189 mol). The bottom ˜35 L was filtered througha pad of Celite and then the phase break was made. The aqueous layer wasre-extracted with DCM (10 L). The organics were filtered through a padof 3 kg magnesol, washing with DCM (8 L). The filtrate was evaporated,triturated with hexane and filtered.

Despite the in-process assay indicating 97% completion, this initialproduct from all four runs typically contained ˜10% SM. These werecombined and triturated in hexane (2 L per kg material) at 50° C., thencooled to 15° C. and filtered to afford Compound 2a (30.0 kg, ˜95%purity, 149 mol, 67%). Mother liquors from the initial trituration andthe re-purification were chromatographed (20 kg silica, eluent 25-50%EtOAc in hexane) to afford additional Compound 2a (4.7 kg, ˜99% purity,24.4 mol, 11%).

Procedure B:

Alternatively the bromination was performed by employing HOAc instead ofHBr. In one specific example, aminopyridine (952 g, 8.49 mmol) wasdissolved in HOAc (7 L) and was treated with NaOAc (1.04 kgs, 12.7 mmol)followed by the dropwise addition of Br₂ (with a dropping funnel-ice wasused to cool the reaction). After the addition of Br₂ the reaction wasallowed to stir at rt overnight. The reaction mixture was poured intowater and made basic with the addition of 6N NaOH. The reaction wasextracted with EtOAc. A significant amount of solid did not dissolve inthe organic or aqueous phase. The entire mixture was filtered and thephases separated. The organic layer was dried (MgSO₄) filtered over aSiO₂ plug eluting with EtOAc. The filtrate was evaporated to give abrown solid, 889 g.

Procedure C:

Alternatively the bromination was performed by employing H₂SO₄. In onespecific example, to 93% sulfuric acid (12.5 kg, 119 mol) in water (26L) in a 50 L reactor was added 2-amino-5-fluoropyridine (6.5 kg, 58mol). The temperature was adjusted to 30° C. then bromine (10 kg, 63mol) added in ten portions over three hours [exothermic, temperaturekept to 30° C. to 40° C., vent set up to scrub through aq.NaOH/Na₂S₂O₃]. The mixture was stirred at 45° C. for 18 hours, then at50° C. for 5 hours. The mixture was cooled to 15° C. for work-up in a400 L reactor.

Four of the above reactions (4×6.5 kg) were combined and quenched in toa mixture of 50% sodium hydroxide (110 kg, 1375 mol) and sodiumthiosulfate (1.8 kg, 11.4 mol) in water (100 L) at −3° C. over one hour.The temperature was adjusted to 32° C. and the slurry filtered andwashed with water (80 L) to afford water-wet crude product (62 kg). Asecond run of three reactions (3×6.5 kg SM) was similarly carried out toafford water-wet crude product (41 kg). The crude products (103 kg) weredissolved (some insolubles) in toluene (280 kg) at 25° C. to 30° C.Brine (20 kg) was added but phase break not possible due to solids. Themixture was filtered through a pad of Celite, washing with toluene, andthe layers then separated. The organics were concentrated to 347 Lvolume to azeotrope residual water for the use of the preparation ofcompound 4-3a. An aliquot was used to determine product concentration asbeing 181 g per liter of solution. Yield=62.8 kg. An additional 600 gwas isolated by extraction of the water/brine layer with ethyl acetate(10 L), and subsequent filtration through a pad of magnesol, evaporationand trituration with hexane. Total yield is 82%.

Example 2: Preparation of5-fluoro-3-((trimethylsilyl)ethynyl)pyridin-2-amine (4-3a)

Procedure A:

To an inert 400 L reactor was charged 4-2a (27.5 kg, 96% purity, 138mol), Pd(PPh₃)₄ (1044 g, 0.90 mol) and CuI (165 g, 0.87 mol), followedby toluene (90 kg). The mixture was de-oxygenated with threevacuum-nitrogen cycles, then triethylamine (19.0 kg, 188 mol) was added.The mixture was de-oxygenated with one more vacuum-nitrogen cycle, thenTMS-acetylene (16.5 kg, 168 mol) was added. The mixture was heated to48° C. for 23 hours (the initial exotherm took the temperature to 53° C.maximum), then cooled to 18° C. The slurry was filtered through a pad ofCelite and washed with toluene (80 kg). The filtrate was washed with 12%Na₂HPO₄ (75 L), then filtered through a pad of silica (25 kg), washingwith 1:1 hexane:MTBE (120 L). This filtrate was evaporated to a brownoil and then dissolved in NMP for the next step. Weight of a solution ofCompound 4-3a-58 kg, ˜50 wt %, 138 mol, 100%. ¹H NMR (CDCl₃, 300 MHz): δ7.90 (s, 1H); 7.33-7.27 (m, 1H); 4.92 (s, NH₂), 0.28 (s, 9H) ppm.

Procedure B:

2-Amino-3-bromo-5-fluoropyridine (4-2a: 10.7 g, 56 mmol) was treatedwith CuI (1.72 g, 9.03 mmol), Pd (dppf)Cl₂ (2.87 g, 3.92 mmol), TMSacetylene (8.25 g, 11.8 mL, 84 mmol), THF (200 mL) and Et₃N (190 mL) andwarmed to reflux overnight. The reaction was judged complete by TLC andpoured into water (200 mL). Phases separated and the phases wereextracted with EtOAc (3×200 mL). Organic phases were combined and dried(MgSO₄), filtered and filtrate concentrated in vacuo to give an oil thatsolidified on vacuum. The solid was dissolved in CH₂Cl₂ and run througha plug of SiO₂ eluting with CH₂Cl₂ to give a yellow solid, 11.7 g, 93%yield.

Example 3: Preparation of 5-fluoro-1-tosyl-1H-pyrrolo[2,3-b]pyridine(7a)

To an inert 400 L reactor was charged potassium t-butoxide (17.5 kg, 156mol) and NMP (45 kg). The mixture was heated to 54° C. then a solutionof compound 4-3a (29 kg, 138 mol) in NMP (38 kg) was added over 2.75hours and rinsed in with NMP (6 kg) (exothermic, maintained at 70°C.-77° C.). The reaction was stirred at 74° C. for 2 hours then cooledto 30° C. and a solution of tosyl chloride (28.5 kg, 150 mol) in NMP (30kg) added over 1.5 hours and rinsed in with NMP (4 kg). The reaction wasexothermic and maintained at 30° C.-43° C. The reaction was stirred for1 hour while cooling to 20° C. then water (220 L) was added over 35minutes (exothermic, maintained at 18° C.-23° C.). The mixture wasstirred at 20° C. for 30 minutes then filtered and washed with water(100 L). The solids were dissolved off the filter with DCM (250 kg),separated from residual water and the organics filtered through a pad ofmagnesol (15 kg, top) and silica (15 kg, bottom), washing with extra DCM(280 kg). The filtrate was concentrated to a thick slurry (˜50 L volume)then MTBE (30 kg) was added while continuing the distillation atconstant volume (final distillate temperature of 51° C.). AdditionalMTBE (10 kg) was added and the slurry cooled to 15° C., filtered andwashed with MTBE (40 L) to afford Compound 7a (19.13 kg, 95% purity,62.6 mol, 45%). Partial concentration of the filtrate afforded a secondcrop (2.55 kg, 91% purity, 8.0 mol, 6%). ¹H NMR (CDCl₃, 300 MHz): δ8.28-8.27 (m, 1H); 8.06-8.02 (m, 2H); 7.77 (d, J=4.0 Hz, 1H); 7.54-7.50(m, 1H); 7.28-7.26 (m, 2H); 6.56 (d, J=4.0 Hz, 1H); 2.37 (s, 3H) ppm.

Example 4: Preparation of3-bromo-5-fluoro-1-tosyl-1H-pyrrolo[2,3-b]pyridine (6a)

Procedure A:

To a slurry of N-bromosuccinimide (14.16 kg, 79.6 mol) in DCM (30 kg) at15° C. was charged a solution of Compound 7a (19.13 kg, 95% purity, and2.86 kg, 91% purity, 71.6 mol) in DCM (115 kg), rinsing in with DCM (20kg). The mixture was stirred at 25° C. for 18 hours, and then cooled to9° C. and quenched by addition of a solution of sodium thiosulfate (400g) and 50% sodium hydroxide (9.1 kg) in water (130 L). The mixture waswarmed to 20° C. and the layers were separated and the organics werewashed with 12% brine (40 L). The aqueous layers were sequentiallyre-extracted with DCM (4×50 kg). The organics were combined and 40 Ldistilled to azeotrope water, then the solution was filtered through apad of silica (15 kg, bottom) and magensol (15 kg, top), washing withDCM (180 kg). The filtrate was concentrated to a thick slurry (˜32 Lvolume) then hexane (15 kg) was added. Additional hexane (15 kg) wasadded while continuing the distillation at constant volume (finaldistillate temperature 52° C.). The slurry was cooled to 16° C.,filtered and washed with hexane (25 kg) to afford Compound 6a (25.6 kg,69.3 mol, 97%). ¹H NMR (CDCl₃, 300 MHz): δ 8.34-8.33 (m, 1H); 8.07 (d,J=8.2 Hz, 2H); 7.85 (s, 1H); 7.52-7.49 (m, 1H); 7.32-7.28 (m, 2H); 2.40(s, 3H) ppm.

Procedure B.

A solution of Br₂ (115 mL, 1.15 eq) in CH₂Cl₂ (1 L) was added, dropwise,to a solution of compound 7a (566 g, 1.95 mol) in CH₂Cl₂ (4 L) over 90minutes. During the addition the temperature increased from 16° C. to23° C. and the reaction mixture was cooled with an ice-salt bath to 10°C. After the addition was complete the temperature had reached 12° C.The suspension (an orange solid had formed during addition) was stirredfor 30 minutes. The reaction mixture was stirred at RT overnight. Sat.aq. NaHCO₃ (4 L) was added, carefully, over 5-10 minutes. The reactionmixture was stirred vigorously for 1 hour and the layers were allowed toseparate. The resulting solution was filtered over a filter. The organiclayer was washed with sat. aq. NaHCO₃ (2 L) and brine (2×1 L), driedover Na₂SO₄ and flushed over silica (2 kg), eluting with CH₂Cl₂ (˜10 Ltotal). The solvents (˜20 L) were removed under reduced pressure to givecompound 6a (580 g) as a white solid. The product was redissolved inCH₂Cl₂ (2.5 L) and filtered over another filter with silica (2 kg),eluting with CH₂Cl₂. After the solvents were removed under reducedpressure compound 7a (568 g, 79% yield) was obtained as an off-whitesolid. After a test reaction for the next step the remaining materialwas washed with heptanes (2×) and dried to give better results in thenext step. ¹H NMR (CDCl₃, 300 MHz): δ 8.34-8.33 (m, 1H); 8.07 (d, J=8.2Hz, 2H); 7.85 (s, 1H); 7.52-7.49 (m, 1H); 7.32-7.28 (m, 2H); 2.40 (s,3H) ppm.

Example 5: Preparation of3-(1,5-dimethyl-2,4-dioxa-3-borabicyclo[3.1.0]hexan-3-yl)-5-fluoro-1-tosyl-1H-pyrrolo[2,3-b]pyridine (1a)

To an inert 400-L reactor was charged Compound 6a (25.6 kg, 69.3 mol),bis(pinacolato)diboron (19 kg, 74.8 mol), potassium acetate (19 kg, 194mol), palladium acetate (156 g, 0.69 mol) and triphenylphosphine (564 g,2.15 mol), followed by dioxane (172 kg), that had been separatelyde-oxygenated using vacuum-nitrogen cycles (×3). The mixture was stirredand de-oxygenated using vacuum-nitrogen cycles (×2), then heated to 100°C. for 15 hours. The mixture was cooled to 35° C. then filtered, washingwith 30° C. THF (75 kg). The filtrate was evaporated and the residuedissolved in DCM (˜90 L). The solution was stirred with 1 kg carbon and2 kg magnesol for 45 minutes then filtered through a pad of silica (22kg, bottom) and magensol (10 kg, top), washing with DCM (160 kg). Thefiltrate was concentrated to a thick slurry (˜40 L volume) thentriturated at 35° C. and hexane (26 kg) was added. The slurry was cooledto 20° C., filtered and washed with a mix of DCM (5.3 kg) and hexane (15kg), then hexane (15 kg) and dried under nitrogen on the filter toafford Compound 1a (23.31 kg, 56.0 mol, 81%) as a white solid. ¹H-NMRconsistent with desired product, HPLC 99.5%, palladium assay 2 ppm. ¹HNMR (CDCl₃, 300 MHz): δ 8.25 (s, 1H); 8.18 (s, 1H); 8.09-8.02 (m, 2H);7.91-7.83 (m, 1H); 7.30-7.23 (m, 2H); 2.39 (s, 3H); 1.38 (s, 12H) ppm.

Example 6: Preparations of 5-fluoro-3-iodopyridin-2-amine (4-2b)

H₂SO₄ (120 mL) was added, dropwise, to a solution of2-amino-5-fluoropyridine (4-1a) (1 kg, 8.9 mol) in AcOH (4 L) and H₂O (1L) over 5 minutes. Periodic acid (H₅IO₆; 450 g, 1.97 mol, 0.22 eq) andI₂ (1 kg, 3.94 mol, 0.44 eq) were added and the reaction mixture wasstirred at 82° C. (internal) overnight. A sample (diluted with H₂O, madealkaline with 30% NaOH, extracted with EtOAc, conc.) showed 13-15%starting material. More H₅IO₆ (80 g) and I₂ (180 g) were added andstirring was continued at 80° C. overnight. The external heating wasremoved and the reaction mixture was stirred at RT overnight. Thereaction mixture was poured over ice-water (8 L), made alkaline with 33%aq. NaOH (˜6.5 L needed) and stirred for 2 h. The precipitated productwas collected by filtration, and washed with hot H₂O (8×3 L). The filterwash left overnight after which the product was washed with heptanes(3×). The product was dried in the stove at 45° C. over the weekend.Compound 2b (1390 g, 65% yield) was obtained as a black solid. H₂O wasadded to the heptanes layer and it was left over the weekend. The darkaqueous layer was separated from the light-yellow organic layer, whichwas concentrated to dryness. More compound 2b (95 g, 70% total yield)was thus obtained as a yellow solid. ¹H NMR (DMSO-d₆, 300 MHz): δ7.95-7.88 (m, 2H) ppm. ¹H NMR (CDCl₃, 300 MHz): δ7.95-7.90 (m, 1H);7.68-7.62 (m, 1H); 4.85 (s, NH₂) ppm.

Example 7: Preparations of 5-fluoro-3-iodopyridin-2-amine (4-3a)

A solution of compound 4-2b (790 g, 3.3 mol) in THF (2.9 L) was degassed(3×), using N₂ (g)/vacuum cycles. Purging with N₂ (g) was startedfollowed by addition of CuI (6.32 g, 0.01 eq), PdCl₂(PPh₃)₂ (23.4 g,0.01 eq) and Et₃N (1.4 L, 3 eq). Purging was continued for 10 minutesand the reaction mixture was degassed once, followed by dropwiseaddition of trimethylsilylacetylene (605 mL, 1.3 eq) over 40-45 minutes.During addition the exothermic reaction did not start by itself and thereaction was heated to ˜45° C. The external heating was removed. Theexothermic reaction had started by this time and the temperature reached˜66° C. (40 minutes after the addition was finished). The reactionmixture was allowed to stir for another 2 h after which the temperaturehad lowered to 26° C. A sample (filtered over Celite, conc.) showedcomplete conversion and the reaction mixture was diluted with EtOAc (3L). The solution was filtered over silica (2 Kg), eluting with EtOAc (9L total). The solvents were removed under reduced pressure to givecompound 4-3a (642 g, 93% yield) as a dark oil. ¹H NMR (CDCl₃, 300 MHz):δ 7.90 (s, 1H); 7.33-7.27 (m, 1H); 4.92 (s, NH₂), 0.28 (s, 9H) ppm.

Example 8: Preparations of tosylate salt of(R)-3-((5-fluoro-2-(5-fluoro-1-tosyl-1H-pyrrolo[2,3-b]pyridin-3-yl)pyrimidin-4-yl)amino)-4,4-dimethylpentanoicAcid (3b-TsOH)

The Suzuki coupling was performed under a N₂ atmosphere by first takingup the chloropyrimidine 2a (1.73 kg) and boronic ester 1a (3.26 kg) in 9vol CH₃CN and 1 vol water. The stiff slurry was sparged with N₂ for 30minutes followed by addition of 0.5 mol % PdCl₂(Amphos)₂ as catalyst.Hunig's base (3 eq.) was added over 30 minutes and the now thin slurryheated to 71° C. overnight. The reaction was judged complete at 99.3%AUC and the mixture cooled to 20° C. Celite (590 g) was slurried withthe reaction mixture for 1 h and then passed through a pad of Celite toremove most of the Pd. The cake was washed with i-PrOAc and the solutionsolvent switched to 6 volumes of i-PrOAc. A 5 wt % aqueous solution ofNaCl (3 vol) was added and the mixture adjusted to pH 5 with of 6N HCl.The aqueous layer was drained to waste and the organic layer treatedwith 1.21 kg of MP-TMT (35 wt % based on theoretical product amount)overnight. With the Pd level brought below <1 ppm, the resin was removedby filtration. The filtration was slow so the mixture was diluted with4.3 L of i-PrOAc. After the volume of the mixture was brought to 8volumes by vacuum distillation, a TsOH solution in 1.4 volumes of2-MeTHF was added to give stiff slurry. TBME (20 vol) was added over 1hour at 20 to 25° C. and the slurry stirred overnight. The solids werecollected by suction filtration and dried on the funnel for 2 days togive 4240 g (97% yield; 1 ppm Pd; 99.31% AUC, Compound (Ia)-0.31% AUC,Compound (2a)-0.12% AUC, RRT 1.01-2.08% AUC, RRT 1.06-0.36% AUC) of3b-TsOH.

Example 8: Preparations of((R)-3-((5-fluoro-2-(5-fluoro-1H-pyrrolo[2,3-b]pyridin-3-yl)pyrimidin-4-yl)amino)-4,4-dimethylpentanoicAcid (Ia)

3b-TsOH (4.14 kg) was taken up in 6 vol of 2-MeTHF as a slurry to which5 eq. of LiOH in 4 vol water were added over 15 minutes at 20° C. to 25°C. The solution was heated to 53° C. overnight to full conversion. Aftercooling the mixture to 23° C., the pH was adjusted to 5.5 to 6 with 6NHCl, the layers separated, and the aqueous layer extracted with 2 vol of2-MeTHF. The combined organic layers were washed twice with 3.6 vol ofpH 6 potassium phosphate buffer. The organic solution was concentratedby vacuum distillation at 40° C. to 4 vol (16 L). Heptane (8 vol, 33 L)was added and the slurry cooled to 20° C. over 5 h and held overnight.The solids were collected by suction filtration and the reactor and cakedried on the filter overnight to give 2068 g (93% yield; 0 ppm residualTsOH; 99.6% AUC, RRT 1.17-0.36% AUC, RRT 0.74-<0.05% AUC) of crude Iafree form.

Example 9: Catalyst Screening for Preparation of Compound Ia

Catalysts, as described in Tables 1 and 2, were screened for activity inSuzuki cross coupling reactions involving compounds 1a and 2a.

TABLE 1 Suzuki screening reactions conducted at 75° C.-80° C. Addi-prod. Catalyst Ligand Base Solvent^(a) tive 3b Pd-132 — K₂CO₃ DMF/H₂O —— Pd-132 — Cs₂CO₃ DMF/H₂O — — Pd-132 — Na₂CO₃ IPA/H₂O — 33% Pd-132 —K₂CO₃ IPA/H₂O — 28% Pd-132 — Cs₂CO₃ IPA/H₂O —  2% Pd-132 — NaO^(t)BuIPA/H₂O —  6% Pd-132 — KO^(t)Bu IPA/H₂O — — Pd-132 — Et₃N PhMe/H₂O —  4%Pd-132 — Et₃N DME/H₂O —  1% Pd-132 — Et₃N CH₃CN/H₂O — 82% Pd-132 — Et₃NDMF/H₂O — 11% Pd-132 — Et₃N IPA/H₂O — 75% Pd-132 — DIPEA PhMe — — Pd-132— DIPEA PhMe/H₂O —  5% Pd-132 — DIPEA THF — — Pd-132 — DIPEA THF/H₂O —61% Pd-132 — DIPEA DME — — Pd-132 — DIPEA DME/H₂O — 54% Pd-132 — DIPEACH₃CN/H₂O — 86% Pd-132 — DIPEA DMF —  2% Pd-132 — DIPEA DMF/H₂O — 80%Pd-132 — DIPEA IPA/H₂O — 64% Pd-132 — K₃PO₄ PhMe/H₂O —  1% Pd-132 —K₃PO₄ THF/H₂O —  6% Pd-132 — K₃PO₄ CH₃CN/H₂O —  3% Pd-132 — K₃PO₄DMF/H₂O — 24% Pd(OAc)₂ Ph₃P K₃PO₄ CH₃CN/H₂O — 48% Pd(OAc)₂ Ph₃P HunigCH₃CN/H₂O — 56% Pd(OAc)₂ Ph₃P K₃PO₄ THF/H₂O —  3% Pd(OAc)₂ Ph₃P HunigTHF/H₂O — 12% Pd(OAc)₂ X-Phos K₂CO₃ THF/H₂O — 79% Pd(OAc)₂ X-Phos DIPEATHF/H₂O — 66% Pd(OAc)₂ X-Phos K₂CO₃ CH₃CN/H₂O — 47% Pd(OAc)₂ X-PhosDIPEA CH₃CN/H₂O — 42% PdCl₂(dppf) — K₃PO₄ CH₃CN/H₂O — 20% PdCl₂(dppf) —K₂CO₃ CH₃CN/H₂O — 28% PdCl₂(dppf) — DIPEA CH₃CN/H₂O — 27% PdCl₂(Ph₃P)₂ —K₃PO₄ CH₃CN/H₂O — 38% PdCl₂(Ph₃P)₂ — K₂CO₃ CH₃CN/H₂O — 64% PdCl₂(Ph₃P)₂— DIPEA CH₃CN/H₂O — 66% Pd₂(dba)₃ [H^(t)BU₃P]BF₄ K₂CO₃ dioxane/H₂O —  2%Pd₂(dba)₃ [H^(t)BU₃P]BF₄ K₂CO₃ CH₃CN/H₂O —  4% Pd₂(dba)₃ [H^(t)BU₃P]BF₄DIPEA dioxane/H₂O —  0% Pd₂(dba)₃ [H^(t)BU₃P]BF₄ DIPEA CH₃CN/H₂O —  2%

In Table 1, Pd-132 refers to the palladium catalyst PdCl₂(AmPhos)₂having the following structure

And, X-Phos refers to the palladium catalyst ligand2-dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl.

TABLE 2 Suzuki screening reactions conducted at 24° C. with CsFadditive. Addi- prod. Catalyst Ligand Base Solvent^(a) tive 3b Pd₂(dba)₃[H^(t)BU₃P]BF₄ K₃PO₄ THF/H₂O CsF 10% Pd₂(dba)₃ [H^(t)BU₃P]BF₄ DIPEATHF/H₂O CsF 14% Pd₂(dba)₃ [H^(t)BU₃P]BF₄ K₃PO₄ CH₃CN/H₂O CsF 17%Pd₂(dba)₃ [H^(t)BU₃P]BF₄ DIPEA CH₃CN/H₂O CsF 5% Pd-132 — DIPEA CH₃CN/H₂OCsF 1% PdCl₂(^(c)Hex₃P)₂ — DIPEA CH₃CN/H₂O CsF 1% PdCl₂(dppf) — DIPEACH₃CN/H₂O CsF 2% PdCl₂(Ph₃P)₂ — DIPEA CH₃CN/H₂O CsF 18%

Other Embodiments

All publications and patents referred to in this disclosure areincorporated herein by reference to the same extent as if eachindividual publication or patent application were specifically andindividually indicated to be incorporated by reference. Should themeaning of the terms in any of the patents or publications incorporatedby reference conflict with the meaning of the terms used in thisdisclosure, the meaning of the terms in this disclosure are intended tobe controlling. Furthermore, the foregoing discussion discloses anddescribes merely exemplary embodiments of the invention. One skilled inthe art will readily recognize from such discussion and from theaccompanying drawings and claims, that various changes, modificationsand variations can be made therein without departing from the spirit andscope of the invention as defined in the following claims.

What is claimed is:
 1. A process for preparing a compound of Formula I:

or a pharmaceutically acceptable salt thereof, wherein R¹ is —Cl or —F;comprising the step of: i) reacting a compound of Formula 1 with acompound of Formula 2

in the presence of water, an organic solvent, a base, and a transitionmetal catalyst to generate a compound of Formula I; wherein each of R⁸and R⁹ is —OH, —O—C₁₋₄ aliphatic optionally substituted with 1-5occurrences of R¹¹, or R⁸ and R⁹ together with the boron atom to whichthey are attached, form a 5-9 membered mono- or bicyclic ring system,optionally substituted with 1-6 occurrences of R¹¹, or BF₃K; each R¹¹ isindependently selected from halogen, —OCH₃, —OH, —NO₂, —NH₂, —SH, —SCH₃,—NHCH₃, —CN, ═O, or unsubstituted —C₁₋₂ aliphatic; X¹ is —Cl, —Br, —I,—OTs, —OMs, —OH, or —NH₂; and the transition metal catalyst is

or the transition metal catalyst comprises Pd and a ligand comprising


2. The process of claim 1, wherein the base of step i) is an organicbase.
 3. The process of claim 2, wherein the organic base comprises atertiary amine.
 4. The process of claim 3, wherein the tertiary aminecomprises diisopropylethylamine, triethylamine, triethylenediamine, orany combination thereof.
 5. The process of any one of claims 1-4,wherein the organic solvent of step i) is an aprotic solvent.
 6. Theprocess of claim 5, wherein the aprotic solvent is acetonitrile,toluene, N,N-dimethylformamide, N,N-dimethylacetamide, acetone, methyltert-butyl ether, or any combination thereof.
 7. The process of any oneof claims 1-6, wherein R¹ is —F.
 8. The process of any one of claims1-6, wherein R¹ is —Cl.
 9. The process of any one of claims 1-8, whereinX¹ is —Cl.
 10. The process of any one of claims 1-9, wherein thereaction of step i) is performed at a temperature between about 50° C.and about 110° C.
 11. The process of claim 10, wherein the reaction ofstep i) is performed at a temperature between about 60° C. and about 95°C.
 12. The process of claim 11, wherein the reaction of step i) isperformed at a temperature between about 65° C. and about 80° C.
 13. Theprocess of any one of claims 1-12, further comprising the step of: ii)deprotecting a compound of Formula 3 to generate a compound of FormulaI:


14. The process of claim 13, wherein step ii) comprises deprotecting thecompound of Formula 3 in the presence of a base.
 15. The process ofclaim 14, wherein the base comprises an inorganic base.
 16. The processof claim 15, wherein the inorganic base is an alkali metal hydroxide.17. The process of claim 16, wherein the alkali metal hydroxide is LiOH,NaOH, KOH, or any combination thereof.
 18. The process of any one ofclaims 1-17, further comprising the step of: via) reacting a compound ofFormula 8 with a compound of Formula 9,

in the presence of a base and an organic solvent to generate a mixturecomprising a compound of Formula 2 and a compound of Formula 10:


19. The process of claim 18, further comprising the steps of: vii)reacting the mixture comprising the compound of Formula 2 and thecompound of Formula 10 with HCl in the presence of an organic solvent togenerate a mixture of hydrochloride salts of the compound of Formula 2and the compound of Formula 10; and viii) recrystalizing the mixture ofthe hydrochloride salts of the compound of Formula 2 and the compound ofFormula 10 to generate the hydrochloride salt of the compound of Formula2.
 20. The process of any one of claims 1-17, further comprising thesteps of: vib) reacting a compound of Formula 8 with a compound ofFormula 9 in the presence of a solvent and a base to generate thecompound Formula 2


21. The process of claim 20, wherein the base of step vib) is aninorganic base selected from tripotassium phosphate, dipotassiumhydrogen phosphate, dipotassium carbonate, disodium carbonate, trisodiumphosphate, disodium hydrogen phosphate, or any combination thereof. 22.The process of either of claims 20 or 21, wherein the solvent of stepvib) comprises water.
 23. The process of claim 22, wherein the solventof step vib) further comprises an alcohol selected from methanol,ethanol, propanol, iso-propanol, butanol, tert-butanol, or anycombination thereof.
 24. The process of any one of claims 20-23, whereinthe reaction of step vib) is performed at a temperature of from about50° C. to about 100° C.
 25. The process of claim 24, wherein thereaction of step vib) is performed at a temperature of from about 60° C.to about 80° C.
 26. The process of any one of claims 1-25, wherein thecompound of Formula 1 is 1a


27. The process of any one of claims 1-26, wherein the compound ofFormula 2 is 2a


28. The process of any one of claims 1-27, wherein the compound ofFormula I is Ia